U.S. patent application number 11/555555 was filed with the patent office on 2007-08-23 for rnai inhibition of influenza virus replication.
This patent application is currently assigned to ALNYLAM PHARMACEUTICALS, INC.. Invention is credited to Antonin de Fougerolles, Anke Geick, Rachel Meyers, Tatiana Novobrantseva, Pamela Tan.
Application Number | 20070197460 11/555555 |
Document ID | / |
Family ID | 38006493 |
Filed Date | 2007-08-23 |
United States Patent
Application |
20070197460 |
Kind Code |
A1 |
Fougerolles; Antonin de ; et
al. |
August 23, 2007 |
RNAI INHIBITION OF INFLUENZA VIRUS REPLICATION
Abstract
The invention relates to compositions and methods for modulating
the expression of influenza viral genes, and more particularly to
the downregulation of influenza viral genes by chemically modified
oligonucleotides.
Inventors: |
Fougerolles; Antonin de;
(Brookline, MA) ; Novobrantseva; Tatiana; (Quincy,
MA) ; Tan; Pamela; (Kulmbach, DE) ; Geick;
Anke; (Bayreuth, DE) ; Meyers; Rachel;
(Newton, MA) |
Correspondence
Address: |
FISH & RICHARDSON PC
P.O. BOX 1022
MINNEAPOLIS
MN
55440-1022
US
|
Assignee: |
ALNYLAM PHARMACEUTICALS,
INC.
300 Third Street
Cambridge
MA
02142
|
Family ID: |
38006493 |
Appl. No.: |
11/555555 |
Filed: |
November 1, 2006 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
60732243 |
Nov 1, 2005 |
|
|
|
60748317 |
Dec 7, 2005 |
|
|
|
60799000 |
May 9, 2006 |
|
|
|
Current U.S.
Class: |
514/44A ;
435/456; 435/5 |
Current CPC
Class: |
C12N 2310/14 20130101;
C12N 2760/16111 20130101; A61P 31/16 20180101; C12N 2310/331
20130101; A61P 43/00 20180101; C12N 15/1131 20130101 |
Class at
Publication: |
514/044 ;
435/005; 435/456 |
International
Class: |
A61K 48/00 20060101
A61K048/00; C12Q 1/70 20060101 C12Q001/70; C12N 15/86 20060101
C12N015/86 |
Claims
1. An iRNA agent comprising a sense strand, wherein the sense
strand comprises at least 15 contiguous nucleotides that differ by
no more than 1, 2, or 3 nucleotides from the sense strand sequences
of any one of the agents provided in Tables 1A-1H, agents numbered
AL-DP-2241-AL-DP-8631, and an antisense strand, wherein the
antisense strand comprises at least 15 contiguous nucleotides that
differ by no more than 1, 2, or 3 nucleotides from the antisense
sequences of any one of the agents provided in Tables 1A-1H, agents
numbered AL-DP-2241-AL-DP-8631.
2. An iRNA agent including a sense strand, wherein the sense strand
comprises at least 15 contiguous nucleotides that differ by no more
than 1, 2, or 3 nucleotides from the sense strand sequences of any
one of the agents provided in Tables 1A-1H, agents numbered
AL-DP-2241-AL-DP-8631, and an antisense strand wherein the
antisense strand comprises at least 15 contiguous nucleotides of
the antisense sequences of any one of the agents provided in Tables
1A-1H, agents numbered AL-DP-2241-AL-DP-8631, and wherein the iRNA
agent reduces the expression of its respective target gene in Cos-7
cells engineered to express the respective target gene by more than
20%, 30%, 40%, 50%, 60%, 70%, or 80% compared to cells which have
not been incubated with the iRNA agent.
3. An iRNA agent comprising a sense strand and an antisense strand
each comprising a sequence of at least 16, 17 or 18 nucleotides
which is essentially identical to one of the sequences of any one
of the agents provided in Tables 1A-1H, agents numbered
AL-DP-2241-AL-DP-8631, except that not more than 1, 2 or 3
nucleotides per strand, respectively, have been substituted by
other nucleotides (e.g. adenosine replaced by uracil), while
essentially retaining the ability to reduce the amount of influenza
A plaques formed in cells in a plaque forming assay.
4. The iRNA agent of claim 1, wherein the antisense RNA strand is
30 or fewer nucleotides in length, and the duplex region of the
iRNA agent is 15-30 nucleotide pairs in length.
5. The iRNA agent of any of claim 1, comprising a modification that
causes the iRNA agent to have increased stability in a biological
sample.
6. The iRNA agent of claim 1, comprising a phosphorothioate or a
2'-modified nucleotide.
7. The iRNA agent of claim 1, comprising at least one
5'-uridine-adenine-3' (5'-ua-3') dinucleotide wherein the uridine
is a 2'-modified nucleotide; at least one 5'-uridine-guanine-3'
(5'-ug-3') dinucleotide, wherein the 5'-uridine is a 2'-modified
nucleotide; at least one 5'-cytidine-adenine-3' (5'-ca-3')
dinucleotide, wherein the 5'-cytidine is a 2'-modified nucleotide;
or at least one 5'-uridine-uridine-3' (5'-uu-3') dinucleotide,
wherein the 5'-uridine is a 2'-modified nucleotide.
8. The iRNA agent of claim 6, wherein the 2'-modification is
selected from the group consisting of: 2'-deoxy,
2'-deoxy-2'-fluoro, 2'-O-methyl, 2'-O-methoxyethyl (2'-O-MOE),
2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE),
2'-O-dimethylaminopropyl (2'-O-DMAP),
2'-O-dimethylaminoethyloxyethyl (2'-O-DMAEOE), and
2'-O--N-methylacetamido (2'-O-NMA).
9. The iRNA agent of any of claim 1, comprising a nucleotide
overhang having 1 to 4 unpaired nucleotides.
10. The iRNA agent of claim 9, wherein the nucleotide overhang has
2 or 3 unpaired nucleotides.
11. The iRNA agent of claim 9, wherein the nucleotide overhang is
at the 3'-end of the antisense strand of the iRNA agent.
12. The iRNA agent of claim 1, comprising a cholesterol moiety.
13. The iRNA agent of claim 12, wherein the cholesterol moiety is
conjugated to the 3'-end of the sense strand of the iRNA agent.
14. The iRNA agent of claim 1, wherein the iRNA agent is targeted
for uptake by cells of the lung.
15. The iRNA agent of claim 1, wherein the iRNA agent comprises at
least one non-natural nucleobase.
16. The iRNA agent of claim 15, wherein the non-natural nucleobase
is difluorotolyl, nitroindolyl, nitropyrrolyl, or
nitroimidazolyl.
17. The iRNA agent of claim 15, wherein the non-natural nucleobase
is difluorotolyl.
18. The iRNA agent of claim 15, wherein only one of the two
oligonucleotide strands comprising the double-stranded
oligonucleotide contains a non-natural nucleobase.
19. The iRNA agent of claim 15, wherein both of the oligonucleotide
strands comprising the double-stranded oligonucleotide
independently contain a non-natural nucleobase.
20. A method of treating a human subject having a pathological
process mediated in part by the replication of influenza A virus,
wherein the iRNA agent comprises a sense strand wherein the sense
strand comprises at least 15 contiguous nucleotides that differ by
no more than 1, 2, or 3 nucleotides from the sense strand sequences
any one of the agents provided in Tables 1A-1H, agents numbered
AL-DP-2241-AL-DP-8631, and an antisense strand, wherein the
antisense strand comprises at least 15 contiguous nucleotides that
differ by no more than 1, 2, or 3 nucleotides from the antisense
strand sequences of any one of the agents provided in Tables 1A-1H,
agents numbered AL-DP-2241-AL-DP-8631.
21. The method of claim 20, wherein the iRNA agent is administered
in an amount sufficient to reduce the replication of influenza
virus in a cell or tissue of the subject.
22. The method of claim 20, wherein the subject is a human.
23. A pharmaceutical composition, comprising: a.) an iRNA agent of
claim 1; and b.) a pharmaceutically acceptable carrier.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Application Ser.
No. 60/732,243, filed Nov. 1, 2005; U.S. Ser. No. 60/748,317, filed
Dec. 7, 2005; and U.S. Ser. No. 60/799,000, filed May 9, 2006. The
contents of each of these provisional applications are hereby
incorporated by reference in their entirety.
TECHNICAL FIELD
[0002] The invention relates to the field of influenza viral
therapy and compositions and methods for modulating viral
replication, and more particularly to the down-regulation of a
gene(s) of an influenza virus by oligonucleotides via RNA
interference which are administered locally to the lungs and nasal
passage via inhalation/intranasal administration, or are
administered systemically, e.g. by via intravenous injection.
BACKGROUND
[0003] RNA interference or "RNAi" is a term initially coined by
Fire and co-workers to describe the observation that
double-stranded RNA (dsRNA) can block gene expression when it is
introduced into worms (Fire et al., Nature 391:806-811, 1998).
Short dsRNA directs gene-specific, post-transcriptional silencing
in many organisms, including vertebrates, and has provided a new
tool for studying gene function. This technology has been reviewed
numerous times recently, see, for example Novina, C. D:, and Sharp,
P., Nature 2004, 430:161, and Sandy, P., et al., Biotechniques
2005, 39:215, hereby incorporated by reference.
[0004] Influenza is one of the most widely spread infections
worldwide. It can be deadly: an estimated 20 to 40 million people
died during the 1918 influenza A virus pandemic. In the United
States between 20 and 40 thousand people die from influenza A virus
infection or its complications each year. During epidemics the
number of influenza related hospitalizations may reach over 300,000
in a single winter season.
[0005] Several properties contribute to the epidemiological success
of influenza virus. First, it is spread easily from person to
person by aerosol (droplet infection). Second, small changes in
influenza virus antigens are frequent (antigenic drift) so that the
virus readily escapes protective immunity induced by a previous
exposure to a different variant of the virus. Third, new strains of
influenza virus can be easily generated by reassortment or mixing
of genetic material between different strains (antigenic shift). In
the case of influenza A virus, such mixing can occur between
subtypes or strains that affect different species. The 1918
pandemic is thought to have been caused by a hybrid strain of virus
derived from reassortment between a swine and a human influenza A
virus. At present, there is a spreading concern about the potential
emergence of novel influenza strains infective to humans,
particularly from avian influenza variants, and more particularly
from strain H5N1, by mixing in humans concurrently exposed to human
and avian influenza virus. The close contact between agricultural
birds and their human breeders familiar in most asian societies has
experts convinced that it is not a question of whether but only
when such a mixed strain will arise. A world-wide pandemic could
swiftly ensue, with even graver consequences than in 1918.
[0006] Despite intensive efforts, there is still no effective
therapy for influenza virus infection and existing vaccines are
limited in value in part because of the properties of antigenic
shift and drift described above. For these reasons, global
surveillance of influenza A virus has been underway for many years,
and the National Institutes of Health designates it as one of the
top priority pathogens for biodefense. Although current vaccines
based upon inactivated virus are able to prevent illness in
approximately 70-80% of healthy individuals under age 65, this
percentage is far lower in the elderly or immunocompromised. In
addition, the expense and potential side effects associated with
vaccine administration make this approach less than optimal.
Although the antiviral drugs currently approved in the United
States for treatment and/or prophylaxis of influenza are helpful,
their use is limited due to concerns about side effects,
compliance, and possible emergence of resistant strains.
[0007] US patent application 20040242518 and corresponding WO
04/028471, both filed Sep. 29, 2003, propose a limited number of
RNAi agents for the treatment of influenza. Their efficacy in
humans is not disclosed.
[0008] Therefore, there still remains a need for the development of
effective therapies for the treatment and prevention of influenza
infection in humans and animals, and particularly for therapies
with high efficiency that allow the targeting of a broad range of
influenza subtypes. One prerequisite for high efficiency is that
the active ingredient is not degraded quickly in a physiological
environment.
SUMMARY
[0009] The present invention is based on the in vitro and in vivo
demonstration that influenza virus infection can be inhibited
through intranasal administration of iRNA agents, as well as by
parenteral administration of such agents and the identification of
potent iRNA agents from the MP, NP, PB1, PB2, or PA gene of
influenza virus that can reduce RNA levels of several subtypes of
influenza virus. Based on these findings, the present invention
provides specific compositions and methods that are useful in
reducing influenza virus mRNA levels, influenza virus protein
levels and influenza virus viral titers in a subject, e.g., a
mammal, such as a human.
[0010] The present invention specifically provides iRNA agents
consisting of, consisting essentially of or comprising at least 15
or more contiguous nucleotides of one of the genes of influenza
virus, particularly the MP, NP, PB1, PB2 and PA genes of influenza
virus, and more particularly agents that comprising 15 or more
contiguous nucleotides from one of the sequences provided in Tables
1A-1H. The iRNA agent preferably comprises less than 30 nucleotides
per strand, e.g., 21-23 nucleotides, such as those provided in
Tables 1A-1H. The double stranded iRNA agent can either have blunt
ends or more preferably have overhangs of 1-4 nucleotides from one
or both 3' ends of the agent.
[0011] Further, the iRNA agent can either contain only naturally
occurring ribonucleotide subunits, or can be synthesized so as to
contain one or more modifications to the sugar or base of one or
more of the ribonucleotide subunits that is included in the agent.
The iRNA agent can be further modified so as to be attached to a
ligand that is selected to improve stability, distribution or
cellular uptake of the agent, e.g. cholesterol. The iRNA agents can
further be in isolated form or can be part of a pharmaceutical
composition used for the methods described herein, particularly as
a pharmaceutical composition formulated for delivery to the lungs
or nasal passage or formulated for parental administration. The
pharmaceutical compositions can contain one or more iRNA agents,
and in some embodiments, will contain two or more iRNA agents, each
one directed to a different segment of a influenza virus gene or a
different influenza virus gene.
[0012] One aspect of the present invention relates to a
double-stranded oligonucleotide comprising at least one non-natural
nucleobase. In certain embodiments, the non-natural nucleobase is
difluorotolyl, nitroindolyl, nitropyrrolyl, or nitroimidazolyl. In
a preferred embodiment, the non-natural nucleobase is
difluorotolyl. In certain embodiments, only one of the two
oligonucleotide strands comprising the double-stranded
oligonucleotide contains a non-natural nucleobase. In certain
embodiments, both of the oligonucleotide strands comprising the
double-stranded oligonucleotide independently contain a non-natural
nucleobase.
[0013] The present invention further provides methods for reducing
the level of influenza virus viral RNA in a cell. Such methods
comprise the step of administering one of the iRNA agents of the
present invention to a subject as further described below. The
present methods utilize the cellular mechanisms involved in RNA
interference to selectively degrade the viral RNA in a cell and are
comprised of the step of contacting a cell with one of the
antiviral iRNA agents of the present invention. Such methods can be
performed directly on a cell or can be performed on a mammalian
subject by administering to a subject one of the iRNA
agents/pharmaceutical compositions of the present invention.
Reduction of viral RNA in a cells results in a reduction in the
amount of viral protein produced, and in an organism, results in a
decrease in replicating viral titer (as shown in the Examples).
[0014] The methods and compositions of the invention, e.g., the
methods and iRNA agent compositions can be used with any dosage
and/or formulation described herein, as well as with any route of
administration described herein. Particularly important is the
showing herein of intranasal administration of an iRNA agent and
its ability to inhibit viral replication in respiratory
tissues.
[0015] The details of one or more embodiments of the invention are
set forth in the accompanying drawings and the description below.
Other features, objects, and advantages of the invention will be
apparent from this description, the drawings, and from the claims.
This application incorporates all cited references, patents, and
patent applications by references in their entirety for all
purposes.
BRIEF DESCRIPTION OF DRAWINGS
[0016] FIGS. 1A-1I: Dose-response curves for the inhibition of
target gene expression for selected RNAi agents. The respective
target gene was recombinantly cloned into Cos-7 cells in a plasmid
resulting in expression of an mRNA encoding the target gene and
Renilla luciferase, the cells treated with the RNAi agent, and
Renilla luciferase was quantified. Cells were treated with the RNAi
agent at concentrations of 100 nM, 25 nM, 6.3 nM, 1.6 nM, 400 pM,
100 pM, 24 pM, 6 pM, 1.5 pM, and 380 fM, and IC.sub.50 values
determined by parametrized curve fitting using the program
XLfit.
DETAILED DESCRIPTION
[0017] The term "influenza virus" is used here to refer to any
strain of influenza virus that is capable of causing disease in an
animal or human subject, or that is an interesting candidate for
experimental analysis. Influenza viruses are described in Fields,
B., et al., Fields' Virology, 4.sup.th ed. 2001, Lippincott
Williams and Wilkins; Philadelphia, ISBN: 0781718325. In
particular, the term encompasses any strain of influenza A virus
that is capable of causing disease in an animal or human subject,
or that is an interesting candidate for experimental analysis. A
large number of influenza A isolates have been partially or
completely sequenced. Table 6 presents merely a partial list of
complete sequences for influenza A genome segments that have been
deposited in a public database (The influenza Sequence Database
(ISD), see Macken, C., Lu, H., Goodman, J., & Boykin, L., "The
value of a database in surveillance and vaccine selection." in
Options for the Control of influenza IV. A. D. M. E. Osterhaus, N.
Cox & A. W. Hampson (Eds.) 2001, Elsevier Science, Amsterdam,
pp 103-106). This database also contains complete sequences for
influenza B and C genome segments. The database is available on the
World Wide Web and includes a convenient search engine that allows
the user to search by genome segment, by species infected by the
virus, and by year of isolation. Influenza sequences are also
available on Genbank. Sequences of influenza genes are therefore
readily available to, or determinable by, those of ordinary skill
in the art.
[0018] For ease of exposition the term "nucleotide" or
"ribonucleotide" is sometimes used herein in reference to one or
more monomeric subunits of an RNA agent. It will be understood that
the usage of the term "ribonucleotide" or "nucleotide" herein can,
in the case of a modified RNA or nucleotide surrogate, also refer
to a modified nucleotide, or surrogate replacement moiety, as
further described below, at one or more positions.
[0019] An "RNA agent" as used herein, is an unmodified RNA,
modified RNA, or nucleoside surrogate, each of which is described
herein or is well known in the RNA synthetic art. While numerous
modified RNAs and nucleoside surrogates are described, preferred
examples include those which have greater resistance to nuclease
degradation than do unmodified RNAs. Preferred examples include
those that have a 2' sugar modification, a modification in a single
strand overhang, preferably a 3' single strand overhang, or,
particularly if single stranded, a 5'-modification which includes
one or more phosphate groups or one or more analogs of a phosphate
group.
[0020] An "iRNA agent" (abbreviation for "interfering RNA agent")
as used herein, is an RNA agent, which can downregulate the
expression of a target gene, e.g., influenza virus. While not
wishing to be bound by theory, an iRNA agent may act by one or more
of a number of mechanisms, including post-transcriptional cleavage
of a target mRNA sometimes referred to in the art as RNAi, or
pre-transcriptional or pre-translational mechanisms. An iRNA agent
can be a double stranded iRNA agent.
[0021] A "ds iRNA agent" (abbreviation for "double stranded iRNA
agent"), as used herein, is an iRNA agent which includes more than
one, and preferably two, strands in which interstrand hybridization
can form a region of duplex structure. A "strand" herein refers to
a contigouous sequence of nucleotides (including non-naturally
occurring or modified nucleotides). The two or more strands may be,
or each form a part of, separate molecules, or they may be
covalently interconnected, e.g., by a linker, e.g., a
polyethyleneglycol linker, to form one molecule. At least one
strand can include a region which is sufficiently complementary to
a target RNA. Such strand is termed the "antisense strand." A
second strand of the dsRNA agent, which comprises a region
complementary to the antisense strand, is termed the "sense
strand." However, a ds iRNA agent can also be formed from a single
RNA molecule which is at least partly self-complementary, forming,
e.g., a hairpin or panhandle structure, including a duplex region.
The latter are herein referred to as short hairpin RNAs or shRNAs.
In such case, the term "strand" refers to one of the regions of the
RNA molecule that is complementary to another region of the same
RNA molecule.
[0022] Although, in mammalian cells, long ds iRNA agents can induce
the interferon response which is frequently deleterious, short ds
iRNA agents do not trigger the interferon response, at least not to
an extent that is deleterious to the cell and/or host (Manche et
al., Mol. Cell. Biol. 12:5238, 1992; Lee et al., Virology 199:491,
1994; Castelli et al., J. Exp. Med. 186:967, 1997; Zheng et al, RNA
10:1934, 2004; Heidel et al., "Lack of interferon response in
animals to naked siRNAs" Nature Biotechn. advance online
publication doi:10.1038/nbt1038, Nov. 21, 2004). The iRNA agents of
the present invention include molecules which are sufficiently
short that they do not trigger a deleterious non-specific
interferon response in normal mammalian cells. Thus, the
administration of a composition including an iRNA agent (e.g.,
formulated as described herein) to a subject can be used to
decreased expression of the influenza virus genes in influenza
virus expressing cells in the subject, while circumventing an
interferon response. Molecules that are short enough that they do
not trigger a deleterious interferon response are termed siRNA
agents or siRNAs herein. "siRNA agent" or "siRNA" as used herein,
refers to an iRNA agent, e.g., a ds iRNA agent, that is
sufficiently short that it does not induce a deleterious interferon
response in a mammalian, and particularly a human, cell, e.g., it
has a duplexed region of less than 60 but preferably less than 50,
40, or 30 nucleotide pairs.
[0023] The isolated iRNA agents described herein, including ds iRNA
agents and siRNA agents, can mediate the decreased expression of a
influenza virus nucleic acid, e.g., by RNA degradation. For
convenience, such RNA is also referred to herein as the RNA to be
silenced. Such a nucleic acid is also referred to as a target gene.
Preferably, the RNA to be silenced is a gene product of a influenza
virus gene that is part of an influenzy virus strain that is
pathogenic to humans.
[0024] As used herein, the phrase "mediates RNAi" refers to the
ability of an agent to silence, in a sequence specific manner, a
target gene. "Silencing a target gene" means the process whereby a
cell containing and/or expressing a certain product of the target
gene when not in contact with the agent, will contain and/or
express at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%
less of such gene product when contacted with the agent, as
compared to a similar cell which has not been contacted with the
agent. Such product of the target gene can, for example, be a
messenger RNA (mRNA), a protein, or a regulatory element.
[0025] As used herein, the term "complementary" is used to indicate
a sufficient degree of complementarity such that stable and
specific binding occurs between a compound of the invention and a
target RNA molecule, e.g., a influenza virus mRNA. Specific binding
requires a sufficient degree of complementarity to avoid
non-specific binding of the oligomeric compound to non-target
sequences under conditions in which specific binding is desired,
i.e., under physiological conditions in the case of in vivo assays
or therapeutic treatment, or in the case of in vitro assays, under
conditions in which the assays are performed. The non-target
sequences typically differ from the target sequences by at least 2,
3 or 4 nucleotides.
[0026] As used herein, an iRNA agent is "sufficiently
complementary" to a target RNA, e.g., a target mRNA (e.g., a target
influenza virus mRNA) if the iRNA agent reduces the production of a
protein encoded by the target RNA in a cell. The iRNA agent may
also be "exactly complementary" to the target RNA, e.g., the target
RNA and the iRNA agent anneal, preferably to form a hybrid made
exclusively of Watson-Crick basepairs in the region of exact
complementarity. A "sufficiently complementary" iRNA agent can
include an internal region (e.g., of at least 10 nucleotides) that
is exactly complementary to a target influenza virus RNA. Moreover,
in some embodiments, the iRNA agent specifically discriminates a
single-nucleotide difference. In this case, the iRNA agent only
mediates RNAi if exact complementarity is found in the region
(e.g., within 7 nucleotides of) the single-nucleotide difference.
Preferred iRNA agents will be based on or consist of or comprise
the sense and antisense sequences provided in Table 1A-1H.
[0027] As used herein, "essentially identical" when used referring
to a first nucleotide sequence in comparison to a second nucleotide
sequence means that the first nucleotide sequence is identical to
the second nucleotide sequence except for up to one, two or three
nucleotide substitutions (e.g., adenosine replaced by uracil).
"Essentially retaining the ability to inhibit influenza virus
expression in cultured human influenza virus expressing cells," as
used herein referring to an iRNA agent not identical to but derived
from one of the iRNA agents of Tables 1A-1H by deletion, addition
or substitution of nucleotides, means that the derived iRNA agent
possesses an inhibitory activity not less than 20% of the
inhibitory activity of the iRNA agent of Tables 1A-1H from which it
was derived. For example, an iRNA agent derived from an iRNA agent
of Tables 1A-1H which lowers the amount of influenza virus mRNA
present in cultured human cells infected with influenza virus by
70% may itself lower the amount of influenza virus mRNA present in
cultured human cells infected with influenza virus by at least 50%
in order to be considered as essentially retaining the ability to
inhibit influenza virus replication in cultured human cells
infected with influenza virus. Optionally, an iRNA agent of the
invention may lower the amount of influenza virus mRNA present in
cultured human cells infected with influenza virus by at least
50%.
[0028] As used herein, a "subject" refers to a mammalian organism
undergoing treatment for a disorder mediated by infection with an
influenza virus. The subject can be any mammal, such as a cow,
horse, mouse, rat, dog, pig, goat, or a primate. In the preferred
embodiment, the subject is a human.
[0029] Influenza Viral Characteristics
[0030] Influenza viruses are enveloped, negative-stranded RNA
viruses of the Orthomyxoviridae family. They are classified as
influenza types A, B, and C, of which influenza A is the most
pathogenic and is believed to be the only type able to undergo
reassortment with animal strains. Influenza types A, B, and C can
be distinguished by differences in their nucleoprotein and matrix
proteins. As discussed further below, influenza A subtypes are
defined by variation in their hemagglutinin (HA) and neuraminidase
(NA) genes and usually distinguished by antibodies that bind to the
corresponding proteins.
[0031] The influenza A viral genome consists of ten genes
distributed in eight RNA segments. The genes encode 10 proteins:
the envelope glycoproteins hemagglutinin (HA) and neuraminidase
(NA); matrix protein (referred to as M1 or MP herein);
nucleoprotein (NP); three polymerases (PB1, PB2, and PA) which are
components of an RNA-dependent RNA transcriptase also referred to
as a polymerase or polymerase complex herein; ion channel protein
(M2), and nonstructural proteins (NS1 and NS2). See Julkunen, I.,
et al., Cytokine and Growth Factor Reviews, 12: 171-180, 2001 for
further details regarding the influenza A virus and its molecular
pathogenesis. See also Fields, B., et al., Fields' Virology,
4.sup.th. ed., Philadelphia: Lippincott Williams and Wilkins; ISBN:
0781718325, 2001. The organization of the influenza B viral genome
is extremely similar to that of influenza A whereas the influenza C
viral genome contains seven RNA segments and lacks the NA gene.
[0032] Influenza A virus classification is based on the
hemagglutinin (H1-H15) and neuraminidase (N1-N9) genes. World
Health Organization (WHO) nomenclature defines each virus strain by
its animal host of origin (specified unless human), geographical
origin, strain number, year of isolation, and antigenic description
of HA and NA. For example, A/Puerto Rico/8/34 (H1N1) designates
strain A, isolate 8, that arose in humans in Puerto Rico in 1934
and has antigenic subtypes 1 of HA and NA. As another example,
A/Chicken/Hong Kong/258/97 (H5N1) designates strain A, isolate 258,
that arose in chickens in Hong Kong in 1997 and has antigenic
subtype 5 of HA and 1 of NA. Human epidemics have been caused by
viruses with HA types H1, H2, and H3 and NA types N1 and N2.
[0033] As mentioned above, genetic variation occurs by two primary
mechanisms in influenza virus A. Antigenic drift occurs via point
mutations, which often occur at antigenically significant positions
due to selective pressure from host immune responses, and antigenic
shift (also referred to as reassortment), involving substitution of
a whole viral genome segment of one subtype by another. Many
different types of animal species including humans, swine, birds,
horses, aquatic mammals, and others, may become infected with
influenza A viruses. Some influenza A viruses are restricted to a
particular species and will not normally infect a different
species. However, some influenza A viruses may infect several
different animal species, principally birds (particularly migratory
water fowl), swine, and humans. This capacity is considered to be
responsible for major antigenic shifts in influenza A virus. For
example, suppose a swine becomes infected with an influenza A virus
from a human and at the same time becomes infected with a different
influenza A virus from a duck. When the two different viruses
reproduce in the swine cells, the genes of the human strain and
duck strain may "mix," resulting in a new virus with a unique
combination of RNA segments. This process is called genetic
reassortment. (Note that this type of genetic reassortment is
distinct from the exchange of genetic information that occurs
between chromosomes during meiosis.)
[0034] Like other viruses and certain bacterial species, influenza
viruses replicate intracellularly. Influenza A viruses replicate in
epithelial cells of the upper respiratory tract. However,
monocytes/macrophages and other white blood cells can also be
infected. Numerous other cell types with cell surface glycoproteins
containing sialic acid are susceptible to infection in vitro since
the virus uses these molecules as a receptor.
[0035] Design and Selection of iRNA Agents
[0036] As used herein, "disorders associated with influenza virus
expression" refers to any biological or pathological state that (1)
is mediated at least in part by the presence of an influenza virus
and (2) whose outcome can be affected by reducing the level of the
influenza virus present. Specific disorders associated with
influenza virus expression are noted below.
[0037] The present invention is based on the design, synthesis and
generation of iRNA agents that target viral genes of influenza
virus, and the demonstration of silencing of a viral gene in vitro
in cultured cells after incubation with an iRNA agent, and the
resulting protective effect towards viral infection.
[0038] An iRNA agent can be rationally designed based on sequence
information and desired characteristics. For example, an iRNA agent
can be designed according to the relative melting temperature of
the candidate duplex. Generally, the duplex should have a lower
melting temperature at the 5' end of the antisense strand than at
the 3' end of the antisense strand.
[0039] The present invention provides compositions containing
siRNA(s) and/or shRNA(s) targeted to one or more influenza virus
transcripts. As the description of the influenza virus replicative
cycle presented above demonstrates, various types of viral RNA
transcripts (primary and secondary vRNA, primary and secondary
viral mRNA, and viral cRNA) are present within cells infected with
influenza virus and play important roles in the viral life cycle.
Any of these transcripts are appropriate targets for siRNA mediated
inhibition by either a direct or an indirect mechanism in
accordance with the present invention. siRNAs and shRNAs that
target any viral mRNA transcript will specifically reduce the level
of the transcript itself in a direct manner, i.e., by causing
degradation of the transcript. In addition, as discussed below,
siRNAs and shRNAs that target certain viral transcripts (e.g., MP,
PA, PB1) will indirectly cause reduction in the levels of viral
transcripts to which they are not specifically targeted. In
situations where alternative splicing is possible, as for the mRNA
that encodes MP and M2 and the mRNA that encodes NS1 and NS2, the
unspliced transcript or the spliced transcript may serve as a
target transcript.
[0040] Potential viral transcripts that may serve as a target for
RNAi based therapy according to the present invention include, for
example, 1) any influenza virus genomic segment; 2) transcripts
that encode any viral proteins including transcripts encoding the
proteins PB1, PB2, PA, NP, NS1, NS2, MP, M2, HA, or NA. As will be
appreciated, transcripts may be targeted in their vRNA, cRNA,
and/or mRNA form(s) by a single siRNA or shRNA. However, it may be
that viral mRNA is the sole or primary target of RNAi as suggested
by Ge et al., WO 04/028471.
[0041] For any particular gene target that is selected, the design
of siRNAs or shRNAs for use in accordance with the present
invention will preferably follow certain guidelines. In general, it
is desirable to target sequences that are specific to the virus (as
compared with the host), and that, preferably, are important or
essential for viral function. Although certain viral genes,
particularly those encoding HA and NA are characterized by a high
mutation rate and are capable of tolerating mutations, certain
regions and/or sequences tend to be conserved. According to certain
embodiments of the invention such sequences may be particularly
appropriate targets. As described further below, such conserved
regions can be identified, for example, through review of the
literature and/or comparisons of influenza gene sequences, a large
number of which are publicly available. Also, in many cases, the
agent that is delivered to a cell according to the present
invention may undergo one or more processing steps before becoming
an active suppressing agent (see below for further discussion); in
such cases, those of ordinary skill in the art will appreciate that
the relevant agent will preferably be designed to include sequences
that may be necessary for its processing. One aspect of the present
invention is the recognition that when multiple strains, subtypes,
etc. (referred to collectively as variants), of an infectious agent
exist, whose genomes vary in sequence, it will often be desirable
to select and/or design siRNAs and shRNAs that target regions that
are highly conserved among different variants. In particular, by
comparing a sufficient number of sequences and selecting highly
conserved regions, it will be possible to target multiple variants
with a single siRNA whose duplex portion includes such a highly
conserved region. Generally such regions should be of sufficient
length to include the entire duplex portion of the siRNA (e.g., 19
nucleotides) and, optionally, one or more 3' overhangs, though
regions shorter than the full length of the duplex can also be used
(e.g., 15, 16, 17, or 18 nucleotides). According to certain
embodiments of the invention a region is highly conserved among
multiple variants if it is identical among the variants. According
to certain embodiments of the invention a region (of whatever
length is to be included in the duplex portion of the siRNA, e.g.,
15, 16, 17, 18, or, preferably, 19 nucleotides) is highly conserved
if it differs by at most one nucleotide (i.e., 0 or 1 nucleotide)
among the variants. According to certain embodiments of the
invention such a region is highly conserved among multiple variants
if it differs by at most two nucleotides (i.e., 0, 1, or 2
nucleotides) among the variants. According to certain embodiments
of the invention a region is highly conserved among multiple
variants if it differs by at most three nucleotides or (i.e., 0, 1,
2, or 3 nucleotides) among the variants. According to certain
embodiments of the invention an siRNA includes a duplex portion
that targets a region that is highly conserved among at least 5
variants, at least variants, at least 15 variants, at least 20
variants, at least 25 variants, at least 30 variants, at least 40
variants, or at least 50 or more variants.
[0042] In order to determine whether a region is highly conserved
among a set of multiple variants, the following procedure may be
used. One member of the set of sequences is selected as the base
sequence, i.e., the sequence to which other sequences are to be
compared. Typically the length of the base sequence will be the
length desired for the duplex portion of the siRNA, e.g, 15, 16,
17, 18, or, preferably 19 nucleotides. According to different
embodiments of the invention the base sequence may be either one of
the sequences in the set being compared or may be a consensus
sequence derived, e.g., by determining for each position the most
frequently found nucleotide at that position among the sequences in
the set.
[0043] Having selected a base sequence, the sequence of each member
of the set of multiple variants is compared with the base sequence.
The number of differences between the base sequence and any member
of the set of multiple variants over a region of the sequence is
used to determine whether the base sequence and that member are
highly conserved over the particular region of interest. As noted
above, in various embodiments of the invention if the number of
sequence differences between two regions is either 0; 0 or 1, 0, 1,
or 2; or 0, 1, 2, or 3, the regions are considered highly
conserved. At the positions where differences occur, the siRNA
sequence may be selected to be identical to the base sequence or to
one of the other sequences. Generally the nucleotide present in the
base sequence will be selected. However in certain embodiments of
the invention, particularly if a nucleotide present at a particular
position in a second sequence in the set being compared is found in
more of the sequences being compared than the nucleotide in the
base sequence, then the siRNA sequence may be selected to be
identical to the second sequence. In addition according to certain
embodiments of the invention, if the consensus nucleotide (most
commonly occurring nucleotide) at the position where the difference
occurs is different to that found in the base sequence, the
consensus nucleotide may be used. Note that this may result in a
sequence that is not identical to any of the sequences being
compared (as may the use of a consensus sequence as the base
sequence).
[0044] The inventors have found that a significant proportion of
the sequences selected using the design parameters described
hereinbelow (see Example 1) prove to be efficient in suppressing
viral replication when included in an siRNA or shRNA and tested as
described below.
[0045] Based on the results shown herein, the present invention
provides iRNA agents that reduce influenza virus replication in
cultured cells infected with influenza virus and in a subject, e.g.
a mammalian, for example a human. Tables 1A-1H provide exemplary
iRNA agents targeting influenza virus. Table 1A, C, D, and E list
siRNAs that do not comprise nucleotide modifications except for one
phosphorothioate linkage between the 3'-terminal and the
penultimate thymidines. Table 1B and H list siRNAs wherein all
nucleotides comprising pyrimidine bases are 2'-O-methyl-modified
nucleotides in the sense strand, and all uridines in a sequence
context of 5'-ua-3' as well as all cytidines in a sequence context
of or 5'-ca-3' are 2'-O-methyl-modified nucleotides in the
antisense strand, except for the iRNA agents with duplex identified
AL-DP-2295, AL-DP-2301, and AL-DP-2302, in which all uridines in a
sequence context of 5'-ug-3' are 2'-O-methyl-modified nucleotides
in the antisense strand. These latter siRNAs had no occurrences of
the sequence motifs 5'-ua-3' or 5'-ca-3', and an analysis of
degradation fragments after incubation of these agents in mouse
serum revealed that the sequence motif 5'-ug-3' was the primary
point of endonucleolytic attack.
[0046] Based on these results, the invention specifically provides
an iRNA agent that includes a sense strand having at least 15
contiguous nucleotides of the sense strand sequences of the agents
provided in Tables 1A-1H, and an antisense strand having at least
15 contiguous nucleotides of the antisense sequences of the agents
provided in Tables 1A-1H.
[0047] The iRNA agents shown in Tables 1A-1H are composed of two
strands of 19 nucleotides in length which are complementary or
identical to the target sequence, plus a 3'-TT overhang. The
present invention provides agents that comprise at least 15, or at
least 16, 17, or 18, or 19 contiguous nucleotides from these
sequences. However, while these lengths may potentially be optimal,
the iRNA agents are not meant to be limited to these lengths. The
skilled person is well aware that shorter or longer iRNA agents may
be similarly effective, since, within certain length ranges, the
efficacy is rather a function of the nucleotide sequence than
strand length. For example, Yang, et al., PNAS 99:9942-9947 (2002),
demonstrated similar efficacies for iRNA agents of lengths between
21 and 30 base pairs. Others have shown effective silencing of
genes by iRNA agents down to a length of approx. 15 base pairs
(Byrom, et al., "Inducing RNAi with siRNA Cocktails Generated by
RNase III" Tech Notes 10(1), Ambion, Inc., Austin, Tex.).
[0048] Therefore, it is possible and contemplated by the instant
invention to select from the sequences provided in Tables 1A-1H a
partial sequence of between 15 to 19 nucleotides for the generation
of an iRNA agent derived from one of the sequences provided in
Tables 1A-1H. Alternatively, one may add one or several nucleotides
to one of the sequences provided in Tables 1A-1H, or an agent
comprising 15 contiguous nucleotides from one of these agents,
preferably, but not necessarily, in such a fashion that the added
nucleotides are complementary to the respective sequence of the
target gene, e.g., an influenza virus gene. For example, the first
15 nucleotides from one of the agents can be combined with the 8
nucleotides found 5' to these sequence in the influenza virus mRNA
to obtain an agent with 23 nucleotides in the sense and antisense
strands. All such derived iRNA agents are included in the iRNA
agents of the present invention, provided they essentially retain
the ability to inhibit influenza virus replication in cultured
human cells infected with influenza virus. TABLE-US-00001 TABLE 1A
Exemplary iRNA agents for targeting influenza virus having 80%
target coverage (criterium 1, see example 1) and 79.9% target
efficiency (criterium 2, see example 1), and having an off target
score of less than 16.8 (see example 1) ELISA ELISA Plasmid Target
% (MDCK (Vero expres- SEQ SEQ influ- remaining cells), cells),
sion, Duplex Sense strand sequence ID Antisense strand ID enza
infec- % inhi- % inhi- % inhi- identifier (5'-3') NO: sequence
(5'-3') NO gene tivity.sup.1 bition.sup.2 bition.sup.3 bition.sup.4
AL-DP-2241 uggaagcaauggcuuuccuTT 1 aggaaagccauugcuuccaTT 2 PB1 3
-31 AL-DP-2242 ggcaccaaacgaucuuaugTT 3 cauaagaucguuuggugccTT 4 NP
-10 28 63 AL-DP-2243 aggcaccaaacgaucuuauTT 5 auaagaucguuuggugccuTT
6 NP -34 15 51 AL-DE-2244 gcaccaaacgaucuuaugaTT 7
ucauaagaucguuuggugcTT 8 NP <50 -26 74 76 AL-DE-2245
cuucuaaccgaggucgaaaTT 9 uuucgaccucgguuagaagTT 10 MP -20 -72
AL-DP-2246 gucgaaacguacguucucuTT 11 agagaacguacguuucgacTT 12 MP -2
-70 AL-DE-2247 cucaaagccgagaucgcgcTT 13 gcgcgaucucggcuuugagTT 14 MP
-4 -95 AL-DE-2248 uucuaaccgaggucgaaacTT 15 guuucgaccucgguuagaaTT 16
MP -23 -37 AL-DE-2249 ucuaaccgaggucgaaacgTT 17
cguuucgaccucgguuagaTT 18 MP -1 18 AL-DE-2250 ucgaaacguacguucucucTT
19 gagagaacguacguuucgaTT 20 MP -27 15 AL-DE-2251
cgaaacguacguucucucuTT 21 agagagaacguacguuucgTT 22 MP -16 12
AL-DE-2252 aaacguacguucucucuauTT 23 auagagagaacguacguuuTT 24 MP -21
-24 AL-DE-2253 cccccucaaagccgagaucTT 25 gaucucggcuuugagggggTT 26 MP
-14 6 AL-DE-2254 cccucaaagccgagaucgcTT 27 gcgaucucggcuuugagggTT 28
MP -22 -24 AL-DE-2255 ccucaaagccgagaucgcgTT 29
cgcgaucucggcuuugaggTT 30 MP -11 -14 AL-DE-2256
acaagaccaauccugucacTT 31 gugacaggauuggucuuguTT 32 MP 17 -22
AL-DE-2257 agcgaggacugcagcguagTT 33 cuacgcugcaguccucgcuTT 34 MP 1
-124 AL-DE-2258 cgaggacugcagcguagacTT 35 gucuacgcugcaguccucgTT 36
MP 26 -37 AL-DE-2259 uugcacuugauauuguggaTT 37 uccacaauaucaagugcaaTT
38 MP 12 -18 AL-DE-2260 ugcacuugauauuguggauTT 39
auccacaauaucaagugcaTT 40 MP -5 -76 AL-DE-2261 auacgguuugaaaagagggTT
41 cccucuuuucaaaccguauTT 42 MP -1 5 AL-DE-2262
uacgguuugaaaagagggcTT 43 gcccucuuuucaaaccguaTT 44 MP -6 -6
AL-DE-2263 acgguuugaaaagagggccTT 45 ggcccucuuuucaaaccguTT 46 MP 3
-22 AL-DE-2264 cgguuugaaaagagggccuTT 47 aggcccucuuuucaaaccgTT 48 MP
4 1 AL-DE-2265 cuaaccgaggucgaaacguTT 49 acguuucgaccucgguuagTT 50 MP
13 -52 AL-DE-2266 uaaccgaggucgaaacguaTT 51 uacguuucgaccucgguuaTT 52
MP 27 -107 AL-DE-2267 aaccgaggucgaaacguacTT 53
guacguuucgaccucgguuTT 54 MP <25 42 -73 AL-DE-2268
accgaggucgaaacguacgTT 55 cguacguuucgaccucgguTT 56 MP -16 -18
AL-DE-2269 ccgaggucgaaacguacguTT 57 acguacguuucgaccucggTT 58 MP -21
-43 AL-DE-2270 cgaggucgaaacguacguuTT 59 aacguacguuucgaccucgTT 60 MP
-6 -25 AL-DE-2271 gaggucgaaacguacguucTT 61 gaacguacguuucgaccucTT 62
MP -29 -29 AL-DE-2272 aggucgaaacguacguucuTT 63
agaacguacguuucgaccuTT 64 MP -23 -59 AL-DE-2273
ggucgaaacguacguucucTT 65 gagaacguacguuucgaccTT 66 MP -9 -36
AL-DE-2274 gcuaaagacaagaccaaucTT 67 gauuggucuugucuuuagcTT 68 MP -24
-17 AL-DE-2275 aauccugucaccucugacuTT 69 agucagaggugacaggauuTT 70 MP
-33 -36 AL-DE-2276 uccugucaccucugacuaaTT 71 uuagucagaggugacaggaTT
72 MP 30% -20 -10 AL-DE-2277 cacgcucaccgugcccaguTT 73
acugggcacggugagcgugTT 74 MP -38 -32 AL-DE-2278
acgcucaccgugcccagugTT 75 cacugggcacggugagcguTT 76 MP -26 -17
AL-DE-2279 cgcucaccgugcccagugaTT 77 ucacugggcacggugagcgTT 78 MP -36
-7 AL-DE-2280 gcucaccgugcccagugagTT 79 cucacugggcacggugagcTT 80 MP
-37 -34 AL-DE-2281 caccgugcccagugagcgaTT 81 ucgcucacugggcacggugTT
82 MP -46 -52 AL-DE-2282 gagcgaggacugcagcguaTT 83
uacgcugcaguccucgcucTT 84 MP -31 -62 AL-DE-2283
uauuguggauucuugaucgTT 85 cgaucaagaauccacaauaTT 86 MP 45% -52 -26
AL-DE-2284 uuguggauucuugaucgucTT 87 gacgaucaagaauccacaaTT 88 MP 61%
39 -60 AL-DE-2285 uguggauucuugaucgucuTT 89 agacgaucaagaauccacaTT 90
MP 41% 3 -49 AL-DE-2286 guggauucuugaucgucuuTT 91
aagacgaucaagaauccacTT 92 MP 36% 9 -50 AL-DE-2287
ucaaaugcauuuaucgucgTT 93 cgacgauaaaugcauuugaTT 94 MP 38% -17 -11
AL-DE-2288 caaaugcauuuaucgucgcTT 95 gcgacgauaaaugcauuugTT 96 MP 39%
23 -58 .sup.1in vitro plaque forming assay in MCDK cells as
described in Example 3.1; .sup.2in in vitro ELISA assay in MCDK
cells as described in Example 3.2: .sup.3in in vitro ELISA assay in
MCDK cells as described in Example 3.2; .sup.4in in vitro ELISA
assay in MCDK cells as described in Example 3.2; negative values
indicate that target gene expression was enhanced in treated cells
compared to controls
[0049] TABLE-US-00002 TABLE 1B Exemplary iRNA agents for targeting
influenza virus derived from agents listed in Table 1A by
stabilization towards nucleolytic degradation by nucleotide
modifications Corre- Target sponding SEQ SEQ influ- Duplex
unmodified Sense strand sequence ID Antisense strand ID enza
identifier duplex.sup.1 (5'-3') NO: sequence (5'-3') NO: gene
AL-DP-2289 AL-DP-2241 umggaagcmaaumggcmumumumcmcmumTT 97
aggaaagccmauugcuuccmaTT 98 PB1 AL-DP-2290 AL-DP-2242
ggcmacmcmaaacmgaumcmumumaumgTT 99 cmaumaagaucguuuggugccTT 100 NP
AL-DP-2291 AL-DP-2243 aggcmacmcmaaacmgaumcmumumaumTT 101
aumaagaucguuuggugccuTT 102 NP AL-DP-2292 AL-DP-2244
gcmacmcmaaacmgaumcmumumaumgaTT 103 ucmaumaagaucguuuggugcTT 104 NP
AL-DP-2293 AL-DP-2245 cmumumcmumaacmcmgaggumcmgaaaTT 105
uuucgaccucgguumagaagTT 106 MP AL-DP-2294 AL-DP-2246
gumcmgaaacmgumacmgumumcmumcmumTT 107 agagaacgumacguuucgacTT 108 MP
AL-DP-2295 AL-DP-2247 cmumcmaaagcmcmgagaumcmgcmgcmTT 109
gcgcgaucucggcuuumgagTT 110 MP AL-DP-2296 AL-DP-2248
umumcmumaacmcmgaggumcmgaaacmTT 111 guuucgaccucgguumagaaTT 112 MP
AL-DP-2297 AL-DP-2249 umcmumaacmcmgaggumcmgaaacmgTT 113
cguuucgaccucgguumagaTT 114 MP AL-DP-2298 AL-DP-2250
umcmgaaacmgumacmgumumcmumcmumcmTT 115 gagagaacgumacguuucgaTT 116 MP
AL-DP-2299 AL-DP-2251 cmgaaacmgumacmgumumcmumcmumcmumTT 117
agagagaacgumacguuucgTT 118 MP AL-DP-2300 AL-DP-2252
aaacmgumacmgumumcmumcmumcmumaumTT 119 aumagagagaacgumacguuuTT 120
MP AL-DP-2301 AL-DP-2254 cmcmcmumcmaaagcmcmgagaumcmgcmTT 121
gcgaucucggcuuumgagggTT 122 MP AL-DP-2302 AL-DP-2255
cmcmumcmaaagcmcmgagaumcmgcmgTT 123 cgcgaucucggcuuumgaggTT 124 MP
AL-DP-2303 AL-DP-2256 acmaagacmcmaaumcmcmumgumcmacmTT 125
gugacmaggauuggucuuguTT 126 MP AL-DP-2304 AL-DP-2257
agcmgaggacmumgcmagcmgumagTT 127 cumacgcugcmaguccucgcuTT 128 MP
AL-DP-2305 AL-DP-2258 cmgaggacmumgcmagcmgumagacmTT 129
gucumacgcugcmaguccuucgTT 130 MP AL-DP-2306 AL-DP-2259
umumgcmacmumumgaumaumumgumggaTT 131 uccmacmaaumaucmaagugcmaaTT 132
MP AL-DP-2307 AL-DP-2260 umgcmacmumumgaumaumumgumggaumTT 133
auccmacmaaumaucmaagugcmaTT 134 MP AL-DP-2308 AL-DP-2265
cmumaacmcmgaggumcmgaaacmgumTT 135 acguuucgaccucgguumagTT 136 MP
AL-DP-2309 AL-DP-2266 umaacmcmgaggumcmgaaacmgumaTT 137
umacguuucgaccucgguumaTT 138 MP AL-DP-2310 AL-DP-2267
aacmcmgaggumcmgaaacmgumacmTT 139 gumacguuucgaccucgguuTT 140 MP
AL-DP-2311 AL-DP-2268 acmcmgaggumcmgaaacmgumacmgTT 141
cgumacguuucgaccucgguTT 142 MP AL-DP-2312 AL-DP-2269
cmcmgaggumcmgaaacmgumacmgumTT 143 acgumacguuucgaccucggTT 144 MP
.sup.1duplex identifier of siRNA agent of Table 1A having an
identical nucleotide sequence when nucleotide modifications are
disregarded
[0050] TABLE-US-00003 TABLE 1C Additional exemplary iRNA agents for
targeting influenza virus not listed in Table 1A having at least
50% target coverage (criterium 1, see Example 1) and at least 80%
target efficiency (criterium 2, see Example 1), and having an off
target score of less than 16.8 (see Example 1) ELISA ELISA Plasmid
Target % (MDCK (Vero expres- SEQ SEQ influ- remaining cells),
cells), sion, Duplex Sense strand sequence ID Antisense strand ID
enza infec- % inhi- % inhi- % inhi- identifier (5'-3') NO: sequence
(5'-3') NO gene tivity.sup.1 bition.sup.2 bition.sup.3 bition.sup.4
AL-DP-2313 augagucuucuaaccgaggTT 145 ccucgguuagaagacucauTT 146 MP
-29 10 AL-DP-2314 ucuucuaaccgaggucgaaTT 147 uucgaccucgguuagaagaTT
148 MP -51 -53 AL-DP-2315 gucuucuaaccgaggucgaTT 149
ucgaccucgguuagaagacTT 150 MP -39 -74 AL-DP-2316
cuucagaucgaacggucuaTT 151 uagaccguucgaucugaagTT 152 EB1 -63 -34
AL-DP-2317 cagaucgaacggucuaacaTT 153 uguuagaccguucgaucugTT 154 EB1
-50 -33 AL-DP-2318 agaucgaacggucuaacagTT 155 cuguuagaccguucgaucuTT
156 EB1 -43 -13 AL-DP-2319 caaaaugcuauaaguaccaTT 157
ugguacuuauagcauuuugTT 158 EB1 -42 -65 AL-DP-2320
uucagaucgaacggucuaaTT 159 uuagaccguucgaucugaaTT 160 EB1 -50 -51
AL-DP-2321 uaccacauucccuuauacuTT 161 aguauaagggaaugugguaTT 162 EB1
-35 1 AL-DP-2322 ucuuacauaaaucggacagTT 163 cuguccgauuuauguaagaTT
164 EB1 12 0 AL-DP-2323 gucuuacauaaaucggacaTT 165
uguccgauuuauguaagacTT 166 EB1 21 -31 AL-DP-2324
agucuuacauaaaucggacTT 167 guccgauuuauguaagacuTT 168 EB1 20 -6
AL-DP-2325 ccucugaugauuucgcucuTT 169 agagcgaaaucaucagaggTT 170 EB1
35 -36 AL-DP-2326 ggaugucaauccgacuuuaTT 171 uaaagucggauugacauccTT
172 EB1 40 12 AL-DP-2327 uuugagagagaaggguacuTT 173
aguacccuucucucucaaaTT 174 NP 38 -14 33 AL-DP-2328
aggcaacgaacccgaucguTT 175 acgaucggguucguugccuTT 176 NP 36 -44 65
AL-DP-2329 ggcaacgaacccgaucgugTT 177 cacgaucggguucguugccTT 178 NP
31 -55 58 AL-DP-2330 caacgaacccgaucgugccTT 179
ggcacgaucggguucguugTT 180 NP 33 -43 56 AL-DP-2331
gcaacgaacccgaucgugcTT 181 gcacgaucggguucguugcTT 182 NP <75 24 44
67 AL-DP-2332 gaaaaggcaacgaacccgaTT 183 ucggguucguugccuuuucTT 184
NP 32 10 78 AL-DP-2333 ucgagcucucggacgaaaaTT 185
uuuucguccgagagcucgaTT 186 NP <50 18 53 69 AL-DP-2334
aacgaacccgaucgugccuTT 187 aggcacgaucggguucguuTT 188 NP 35 -19 30
AL-DP-2335 uauuucuucggagacaaugTT 189 cauugucuccgaagaaauaTT 190 NP 2
-20 41 AL-DP-2348 ugcaugauaaaggcaguccTT 191 ggacugccuuuaucaugcaTT
192 PB2 -45 -65 4 AL-DP-2349 auggggaugaucggaauauTT 193
auauuccgaucauccccauTT 194 PB2 0 32 55 AL-DP-2350
uggggaugaucggaauauuTT 195 aauauuccgaucauccccaTT 196 PB2 -41 5 52
AL-DP-2351 gaaacgggacucuagcauaTT 197 uaugcuagagucccguuucTT 198 PB2
<25 -33 68 76 AL-DP-2356 agacuuugugcgacaaugcTT 199
gcauugucgcacaaagucuTT 200 PA 19 -22 77 AL-DP-2357
gacuuugugcgacaaugcuTT 201 agcauugucgcacaaagucTT 202 PA -84 13 86
AL-DP-2358 ucuaugggauuccuuucguTT 203 acgaaaggaaucccauagaTT 204 PA
-33 -81 30 AL-DP-2359 cuaugggauuccuuucgucTT 205
gacgaaaggaaucccauagTT 206 PA -59 -109 80 AL-DP-2360
auguggauggauucgaaccTT 207 gguucgaauccauccacauTT 208 PA 1 4 16
AL-DP-2361 uguggauggauucgaaccgTT 209 cgguucgaauccauccacaTT 210 PA
-10 28 -7 AL-DP-2362 guggauggauucgaaccgaTT 211
ucgguucgaauccauccacTT 212 PA <25 -28 59 88 AL-DP-2363
uggauggauucgaaccgaaTT 213 uucgguucgaauccauccaTT 214 PA <25 -7 50
89 AL-DP-2364 ggauggauucgaaccgaacTT 215 guucgguucgaauccauccTT 216
PA <25 55 65 89 AL-DP-2365 gauggauucgaaccgaacgTT 217
cguucgguucgaauccaucTT 218 PA -3 27 60 AL-DP-2366
auggauucgaaccgaacggTT 219 ccguucgguucgaauccauTT 220 PA 32 19 31
AL-DP-2367 uggauucgaaccgaacggcTT 221 gccguucgguucgaauccaTT 222 PA 0
-52 14 AL-DP-2368 aucuccacaacucgaggggTT 223 ccccucgaguuguggagauTT
224 PA 22 0 20 .sup.1in in vitro plaque forming assay in MCDK cells
as described in Example 3.1; .sup.2in in vitro ELISA assay in MCDK
cells as described in Example 3.2: .sup.3in in vitro ELISA assay in
MCDK cells as described in Example 3.2; .sup.4in in vitro ELISA
assay in MCDK cells as described in Example 3.2; negative values
indicate that target gene expression was enhanced in treated cells
compared to controls
[0051] TABLE-US-00004 TABLE 1D Additional exemplary iRNA agents for
targeting influenza virus not listed in Table 1A or C, and having
at least 80% target coverage (criterium 1, see Example 1) and at
least 80% target efficiency (criterium 2, see Example 1) ELISA
ELISA Plasmid Target % (MDCK (Vero expres- SEQ SEQ influ- remaining
cells), cells), sion, Duplex Sense strand sequence ID Antisense
strand ID enza infec- % inhi- % inhi- % inhi- identifier (5'-3')
NO: sequence (5'-3') NO gene tivity.sup.1 bition.sup.2 bition.sup.3
bition.sup.4 AL-DP-7614 uaacaauagagagaaugguTT 225
accauucucucuauuguuaTT 226 NP 17 +80 35 AL-DP-7635
gaaacguacguucucucuaTT 227 uagagagaacguacguuucTT 228 MP 7 19
AL-DP-7636 aacguacguucucucuaucTT 229 gauagagagaacguacguuTT 230 MP 1
24 AL-DP-7637 uggcuaaagacaagaccaaTT 231 uuggucuugucuuuagccaTT 232
MP <25 42 8 AL-DP-7638 ggcuaaagacaagaccaauTT 233
auuggucuugucuuuagccTT 234 MP 19 5 AL-DP-7639 cuaaagacaagaccaauccTT
235 ggauuggucuugucuuuagTT 236 MP 22 24 AL-DP-7640
uaaagacaagaccaauccuTT 237 aggauuggucuugucuuuaTT 238 MP 11 43
AL-DP-7641 aaagacaagaccaauccugTT 239 caggauuggucuugucuuuTT 240 MP
13 33 AL-DP-7642 aagacaagaccaauccuguTT 241 acaggauuggucuugucuuTT
242 MP -12 39 AL-DP-7643 agacaagaccaauccugucTT 243
gacaggauuggucuugucuTT 244 MP 4 5 AL-DP-7644 gacaagaccaauccugucaTT
245 ugacaggauuggucuugucTT 246 MP 3 26 AL-DP-7645
caagaccaauccugucaccTT 247 ggugacaggauuggucuugTT 248 MP 2 2
AL-DP-7646 aagaccaauccugucaccuTT 249 aggugacaggauuggucuuTT 250 MP
12 9 AL-DP-7647 agaccaauccugucaccucTT 251 gaggugacaggauuggucuTT 252
MP 8 -5 AL-DP-7648 gaccaauccugucaccucuTT 253 agaggugacaggauuggucTT
254 MP -27 2 AL-DP-7649 accaauccugucaccucugTT 255
cagaggugacaggauugguTT 256 MP 8 0 AL-DP-7650 ccaauccugucaccucugaTT
257 ucagaggugacaggauuggTT 258 MP -1 11 AL-DP-7651
caauccugucaccucugacTT 259 gucagaggugacaggauugTT 260 MP <50 30 16
AL-DP-7652 auccugucaccucugacuaTT 261 uagucagaggugacaggauTT 262 MP
<50 69 9 AL-DP-7653 uucacgcucaccgugcccaTT 263
ugggcacggugagcgugaaTT 264 MP <75 31 6 AL-DP-7654
ucacgcucaccgugcccagTT 265 cugggcacggugagcgugaTT 266 MP <75 57 12
AL-DP-7655 cucaccgugcccagugagcTT 267 gcucacugggcacggugagTT 268 MP
22 AL-DP-7656 ucaccgugcccagugagcgTT 269 cgcucacugggcacggugaTT 270
MP <75 69 10 AL-DP-7657 accgugcccagugagcgagTT 271
cucgcucacugggcacgguTT 272 MP 1 22 AL-DP-7658 ccgugcccagugagcgaggTT
273 ccucgcucacugggcacggTT 274 MP <75 34 20 AL-DP-7659
cgugcccagugagcgaggaTT 275 uccucgcucacugggcacgTT 276 MP 4 35
AL-DP-7660 gugcccagugagcgaggacTT 277 guccucgcucacugggcacTT 278 MP
<75 33 49 AL-DP-7661 ugcccagugagcgaggacuTT 279
aguccucgcucacugggcaTT 280 MP <50 -10 58 AL-DP-7662
gcccagugagcgaggacugTT 281 caguccucgcucacugggcTT 282 MP -16 -44
AL-DP-7663 cccagugagcgaggacugcTT 283 gcaguccucgcucacugggTT 284 MP
11 -8 AL-DP-7664 ccagugagcgaggacugcaTT 285 ugcaguccucgcucacuggTT
286 MP 24 -20 AL-DP-7665 cagugagcgaggacugcagTT 287
cugcaguccucgcucacugTT 288 MP 7 -24 AL-DP-7666 agugagcgaggacugcagcTT
289 gcugcaguccucgcucacuTT 290 MP <50 42 -22 AL-DP-7667
gugagcgaggacugcagcgTT 291 cgcugcaguccucgcucacTT 292 MP 20 -17
AL-DP-7668 ugagcgaggacugcagcguTT 293 acgcugcaguccucgcucaTT 294 MP
<50 51 -54 AL-DP-7669 gcgaggacugcagcguagaTT 295
ucuacgcugcaguccucgcTT 296 MP <25 32 37 AL-DP-7670
gaggacugcagcguagacgTT 297 cgucuacgcugcaguccucTT 298 MP <75 35 20
AL-DP-7671 gcacuugauauuguggauuTT 299 aauccacaauaucaagugcTT 300 MP
-19 4 AL-DP-7672 cacuugauauuguggauucTT 301 gaauccacaauaucaagugTT
302 MP 12 -10 AL-DP-7673 acuugauauuguggauucuTT 303
agaauccacaauaucaaguTT 304 MP 0 -1 AL-DP-7674 cuugauauuguggauucuuTT
305 aagaauccacaauaucaagTT 306 MP -8 -36 AL-DP-7675
uugauauuguggauucuugTT 307 caagaauccacaauaucaaTT 308 MP <75 2 -27
AL-DP-7676 ugauauuguggauucuugaTT 309 ucaagaauccacaauaucaTT 310 MP
-29 -40 AL-DP-7677 gauauuguggauucuugauTT 311 aucaagaauccacaauaucTT
312 MP -14 -24 AL-DP-7678 auauuguggauucuugaucTT 313
gaucaagaauccacaauauTT 314 MP 24 -22 AL-DP-7679
ggauucuugaucgucuuuuTT 315 aaaagacgaucaagaauccTT 316 MP 3 -34
AL-DP-7684 ccgucaggcccccucaaagTT 317 cuuugagggggccugacggTT 318 MP
-1 23 AL-DP-7685 cgucaggcccccucaaagcTT 319 gcuuugagggggccugacgTT
320 MP -12 36 AL-DP-7686 gucaggcccccucaaagccTT 321
ggcuuugagggggccugacTT 322 MP -4 14 AL-DP-7687 ucaggcccccucaaagccgTT
323 cggcuuugagggggccugaTT 324 MP <75 32 -11 AL-DP-7688
caggcccccucaaagccgaTT 325 ucggcuuugagggggccugTT 326 MP <25 56
-14 AL-DP-7689 aggcccccucaaagccgagTT 327 cucggcuuugagggggccuTT 328
MP <50 83 -21 AL-DP-7690 ggcccccucaaagccgagaTT 329
ucucggcuuugagggggccTT 330 MP <50 89 -29 AL-DP-7691
gcccccucaaagccgagauTT 331 aucucggcuuugagggggcTT 332 MP 8 -21
AL-DP-7692 ccccucaaagccgagaucgTT 333 cgaucucggcuuugaggggTT 334 MP 5
-86 .sup.1in in vitro plaque forming assay in MCDK cells as
described in Example 3.1; .sup.2in in vitro ELISA assay in MCDK
cells as described in Example 3.2: .sup.3in in vitro ELISA assay in
MCDK cells as described in Example 3.2; .sup.4in in vitro ELISA
assay in MCDK cells as described in Example 3.2; negative values
indicate that target gene expression was enhanced in treated cells
compared to controls
[0052] TABLE-US-00005 TABLE 1E Additional exemplary iRNA agents for
targeting influenza virus not listed in Table 1A, C, or D,and
having at least 50% target coverage (criterium 1, see Example 1)
and at least 80% target efficiency (criterium 2, see Example 1)
ELISA ELISA Plasmid Target % (MDCK (Vero expres- SEQ SEQ influ-
remaining cells), cells), sion, Duplex Sense strand sequence ID
Antisense strand ID enza infec- % inhi- % inhi- % inhi- identifier
(5'-3') NO: sequence (5'-3') NO gene tivity.sup.1 bition.sup.2
bition.sup.3 bition.sup.4 AL-DP-7565 cgagagaggcgaagagacaTT 335
ugucucuucgccucucucgTT 336 PA <50 -29 71 59 AL-DP-7566
gaagagacaauugaagaaaTT 337 uuucuucaauugucucuucTT 338 PA <75 -15
61 57 AL-DP-7567 uuuagagccuauguggaugTT 339 cauccacauaggcucuaaaTT
340 PA <75 -64 49 13 AL-DP-7568 uuagagccuauguggauggTT 341
ccauccacauaggcucuaaTT 342 PA <75 -24 60 19 AL-DP-7569
uagagccuauguggauggaTT 343 uccauccacauaggcucuaTT 344 PA <75 -47
67 26 AL-DP-7570 agagccuauguggauggauTT 345 auccauccacauaggcucuTT
346 PA <50 -23 87 72 AL-DP-7571 gagccuauguggauggauuTT 347
aauccauccacauaggcucTT 348 PA <25 2 94 84 AL-DP-7572
agccuauguggauggauucTT 349 gaauccauccacauaggcuTT 350 PA <25 -14
82 76 AL-DP-7573 uaugaagcaauugaggaguTT 351 acuccucaauugcuucauaTT
352 PA <75 -58 48 6 AL-DP-7574 augaagcaauugaggagugTT 353
cacuccucaauugcuucauTT 354 PA -69 16 4 AL-DP-7575
ugaagcaauugaggagugcTT 355 gcacuccucaauugcuucaTT 356 PA -65 99 2
AL-DP-7576 gaagcaauugaggagugccTT 357 ggcacuccucaauugcuucTT 358 PA
<25 -23 99 63 AL-DP-7577 aagcaauugaggagugccuTT 359
aggcacuccucaauugcuuTT 360 PA <25 -27 99 60 AL-DP-7578
gaucccuggguuuugcuuaTT 361 uaagcaaaacccagggaucTT 362 PA <50 -50
96 80 AL-DP-7579 aucccuggguuuugcuuaaTT 363 uuaagcaaaacccagggauTT
364 PA <25 -51 98 87 AL-DP-7580 ucccuggguuuugcuuaauTT 365
auuaagcaaaacccagggaTT 366 PA <75 15 95 60 AL-DP-7581
cccuggguuuugcuuaaugTT 367 cauuaagcaaaacccagggTT 368 PA <50 -55
100 75 AL-DP-7582 ccuggguuuugcuuaaugcTT 369 gcauuaagcaaaacccaggTT
370 PA <25 3 74 87 AL-DP-7583 ucuugguucaacuccuuccTT 371
ggaaggaguugaaccaagaTT 372 PA <75 43 14 19 AL-DP-7584
cuugguucaacuccuuccuTT 373 aggaaggaguugaaccaagTT 374 PA -19 22 74
AL-DP-7585 aaacgggacucuagcauacTT 375 guaugcuagagucccguuuTT 376 PB2
6 35 66 AL-DP-7586 aacgggacucuagcauacuTT 377 aguaugcuagagucccguuTT
378 PB2 <25 43 35 58 AL-DP-7587 acgggacucuagcauacuuTT 379
aaguaugcuagagucccguTT 380 PB2 <25 14 52 71 AL-DP-7588
cgggacucuagcauacuuaTT 381 uaaguaugcuagagucccgTT 382 PB2 <25 34
36 72 AL-DP-7589 gggacucuagcauacuuacTT 383 guaaguaugcuagagucccTT
384 PB2 -28 32 69 AL-DP-7590 ggacucuagcauacuuacuTT 385
aguaaguaugcuagaguccTT 386 PB2 -10 36 75 AL-DP-7591
gacucuagcauacuuacugTT 387 caguaaguaugcuagagucTT 388 PB2 -20 22 61
AL-DP-7592 acucuagcauacuuacugaTT 389 ucaguaaguaugcuagaguTT 390 PB2
<25 -25 48 77 AL-DP-7593 cucuagcauacuuacugacTT 391
gucaguaaguaugcuagagTT 392 PB2 <75 -14 50 64 AL-DP-7594
ucuagcauacuuacugacaTT 393 ugucaguaaguaugcuagaTT 394 PB2 -40 9 44
AL-DP-7595 cuagcauacuuacugacagTT 395 cugucaguaaguaugcuagTT 396 PB2
<25 59 48 66 AL-DP-7596 uagcauacuuacugacagcTT 397
gcugucaguaaguaugcuaTT 398 PB2 -33 -14 44 AL-DP-7597
agcauacuuacugacagccTT 399 ggcugucaguaaguaugcuTT 400 PB2 -13 -27 29
AL-DP-7598 gcauacuuacugacagccaTT 401 uggcugucaguaaguaugcTT 402 PB2
<25 8 65 73 AL-DP-7599 cauacuuacugacagccagTT 403
cuggcugucaguaaguaugTT 404 PB2 <25 -39 43 69 AL-DP-7600
auacuuacugacagccagaTT 405 ucuggcugucaguaaguauTT 406 PB2 <25 -33
48 74 AL-DP-7601 uacuuacugacagccagacTT 407 gucuggcugucaguaaguaTT
408 PB2 -28 -40 35 AL-DP-7602 acuuacugacagccagacaTT 409
ugucuggcugucaguaaguTT 410 PB2 6 17 60 AL-DP-7603
cuuacugacagccagacagTT 411 cugucuggcugucaguaagTT 412 PB2 <25 16
73 77 AL-DP-7604 uuacugacagccagacagcTT 413 gcugucuggcugucaguaaTT
414 PB2 18 -26 37 AL-DP-7605 uacugacagccagacagcgTT 415
cgcugucuggcugucaguaTT 416 PB2 20 -133 41 AL-DP-7606
acugacagccagacagcgaTT 417 ucgcugucuggcugucaguTT 418 PB2 <25 34 0
80 AL-DP-7607 cugacagccagacagcgacTT 419 gucgcugucuggcugucagTT 420
PB2 <25 38 51 74 AL-DP-7608 ugacagccagacagcgaccTT 421
ggucgcugucuggcugucaTT 422 PB2 <25 64 -155 34 AL-DP-7609
gacagccagacagcgaccaTT 423 uggucgcugucuggcugucTT 424 PB2 <25 88
-16 71 AL-DP-7610 acagccagacagcgaccaaTT 425 uuggucgcugucuggcuguTT
426 PB2 <25 55 29 72 AL-DP-7611 cagccagacagcgaccaaaTT 427
uuuggucgcugucuggcugTT 428 PB2 <25 33 88 78 AL-DP-7612
agccagacagcgaccaaaaTT 429 uuuuggucgcugucuggcuTT 430 PB2 <25 37
73 74 AL-DP-7613 gccagacagcgaccaaaagTT 431 cuuuuggucgcugucuggcTT
432 PB2 <25 57 -4 79 AL-DP-7615 cccgaucgugccuuccuuuTT 433
aaaggaaggcacgaucgggTT 434 NP 17 -80 35 AL-DP-7616
ccgaucgugccuuccuuugTT 435 caaaggaaggcacgaucggTT 436 NP 11 -40 47
AL-DP-7617 cgaucgugccuuccuuugaTT 437 ucaaaggaaggcacgaucgTT 438 NP
22 -81 47 AL-DP-7618 gaucgugccuuccuuugacTT 439
gucaaaggaaggcacgaucTT 440 NP <25 87 86 73 AL-DP-7619
aucgugccuuccuuugacaTT 441 ugucaaaggaaggcacgauTT 442 NP <25 86 82
50 AL-DP-7620 ucgugccuuccuuugacauTT 443 augucaaaggaaggcacgaTT 444
NP <25 3 43 53 AL-DP-7621 cgugccuuccuuugacaugTT 445
caugucaaaggaaggcacgTT 446 NP -22 7 47 AL-DP-7622
gugccuuccuuugacaugaTT 447 ucaugucaaaggaaggcacTT 448 NP <25 17 78
53 AL-DP-7623 ggaucuuauuucuucggagTT 449 cuccgaagaaauaagauccTT 450
NP <25 66 95 58 AL-DP-7624 gaucuuauuucuucggagaTT 451
ucuccgaagaaauaagaucTT 452 NP <25 95 96 75 AL-DP-7625
aucuuauuucuucggagacTT 453 gucuccgaagaaauaagauTT 454 NP <25 33 78
77 AL-DP-7626 ucuuauuucuucggagacaTT 455 ugucuccgaagaaauaagaTT 456
NP 8 17 7 AL-DP-7627 cuuauuucuucggagacaaTT 457
uugucuccgaagaaauaagTT 458 NP <25 32 66 54 AL-DP-7628
uuauuucuucggagacaauTT 459 auugucuccgaagaaauaaTT 460 NP <25 24 81
72 AL-DP-7629 auuucuucggagacaaugcTT 461 gcauugucuccgaagaaauTT 462
NP <50 11 44 61 AL-DP-7630 gggcggggagucuucgagcTT 463
gcucgaagacuccccgcccTT 464 NP 15 24 9 AL-DP-7631
ggcggggagucuucgagcuTT 465 agcucgaagacuccccgccTT 466 NP 5 33 30
AL-DP-7632 gcggggagucuucgagcucTT 467 gagcucgaagacuccccgcTT 468 NP
<25 16 41 33 AL-DP-7633 cggggagucuucgagcucuTT 469
agagcucgaagacuccccgTT 470 NP <25 33 55 50 AL-DP-7634
ggggagucuucgagcucucTT 471 gagagcucgaagacuccccTT 472 NP <25 41 82
44 AL-DP-7680 ugacgauggucauuuugucTT 473 gacaaaaugaccaucgucaTT 474
MP <25 58 72 49 AL-DP-7681 gacgauggucauuuugucaTT 475
ugacaaaaugaccaucgucTT 476 MP -1 23 AL-DP-7682 ucccgucaggcccccucaaTT
477 uugagggggccugacgggaTT 478 MP -12 36 AL-DP-7683
cccgucaggcccccucaaaTT 479 uuugagggggccugacgggTT 480 MP -4 14
AL-DP-8102 ugagucuucuaaccgagguTT 481 accucgguuagaagacucaTT 482 MP
AL-DP-8103 gagucuucuaaccgaggucTT 483 gaccucgguuagaagacucTT 484 MP
AL-DP-8104 agucuucuaaccgaggucgTT 485 cgaccucgguuagaagacuTT 486 MP
AL-DP-8107 auuguggauucuugaucguTT 487 acgaucaagaauccacaauTT 488 MP
AL-DP-8108 uggauucuugaucgucuuuTT 489 aaagacgaucaagaauccaTT 490 MP
AL-DP-8109 gauucuugaucgucuuuucTT 491 gaaaagacgaucaagaaucTT 492 MP
AL-DP-8110 auucuugaucgucuuuucuTT 493 agaaaagacgaucaagaauTT 494 MP
AL-DP-8111 uucuugaucgucuuuucuuTT 495 aagaaaagacgaucaagaaTT 496 MP
AL-DP-8112 ucuugaucgucuuuucuucTT 497 gaagaaaagacgaucaagaTT 498 MP
AL-DP-8113 cuugaucgucuuuucuucaTT 499 ugaagaaaagacgaucaagTT 500
MP
AL-DP-8114 uugaucgucuuuucuucaaTT 501 uugaagaaaagacgaucaaTT 502 MP
AL-DP-8115 ugaucgucuuuucuucaaaTT 503 uuugaagaaaagacgaucaTT 504 MP
AL-DP-8116 gaucgucuuuucuucaaauTT 505 auuugaagaaaagacgaucTT 506 MP
AL-DP-8117 aucgucuuuucuucaaaugTT 507 cauuugaagaaaagacgauTT 508 MP
AL-DP-8118 ucgucuuuucuucaaaugcTT 509 gcauuugaagaaaagacgaTT 510 MP
AL-DP-8119 cgucuuuucuucaaaugcaTT 511 ugcauuugaagaaaagacgTT 512 MP
AL-DP-8120 gucuuuucuucaaaugcauTT 513 augcauuugaagaaaagacTT 514 MP
AL-DP-8121 ucuuuucuucaaaugcauuTT 515 aaugcauuugaagaaaagaTT 516 MP
AL-DP-8122 cuuuucuucaaaugcauuuTT 517 aaaugcauuugaagaaaagTT 518 MP
AL-DP-8123 uuuucuucaaaugcauuuaTT 519 uaaaugcauuugaagaaaaTT 520 MP
AL-DP-8124 uuucuucaaaugcauuuauTT 521 auaaaugcauuugaagaaaTT 522 MP
AL-DP-8125 uucuucaaaugcauuuaucTT 523 gauaaaugcauuugaagaaTT 524 MP
AL-DP-8126 ucuucaaaugcauuuaucgTT 525 cgauaaaugcauuugaagaTT 526 MP
AL-DP-8127 cuucaaaugcauuuaucguTT 527 acgauaaaugcauuugaagTT 528 MP
AL-DP-8128 uucaaaugcauuuaucgucTT 529 gacgauaaaugcauuugaaTT 530 MP
AL-DP-8129 uuaaauacgguuugaaaagTT 531 cuuuucaaaccguauuuaaTT 532 MP
AL-DP-8130 uaaauacgguuugaaaagaTT 533 ucuuuucaaaccguauuuaTT 534 MP
AL-DP-8131 aaauacgguuugaaaagagTT 535 cucuuuucaaaccguauuuTT 536 MP
AL-DP-8132 aauacgguuugaaaagaggTT 537 ccucuuuucaaaccguauuTT 538 MP
AL-DP-8133 uugaaaagagggccuucuaTT 539 uagaaggcccucuuuucaaTT 540 MP
AL-DP-8134 ugaaaagagggccuucuacTT 541 guagaaggcccucuuuucaTT 542 MP
AL-DP-8135 gaaaagagggccuucuacgTT 543 cguagaaggcccucuuuucTT 544 MP
AL-DP-8136 ccugagucuaugagggaagTT 545 cuucccucauagacucaggTT 546 MP
AL-DP-8137 cugagucuaugagggaagaTT 547 ucuucccucauagacucagTT 548 MP
AL-DP-8138 ugcuguggauguugacgauTT 549 aucgucaacauccacagcaTT 550 MP
AL-DP-8139 gcuguggauguugacgaugTT 551 caucgucaacauccacagcTT 552 MP
AL-DP-8140 cuguggauguugacgauggTT 553 ccaucgucaacauccacagTT 554 MP
AL-DP-8141 uguggauguugacgaugguTT 555 accaucgucaacauccacaTT 556 MP
AL-DP-8142 guggauguugacgauggucTT 557 gaccaucgucaacauccacTT 558 MP
AL-DP-8143 uggauguugacgauggucaTT 559 ugaccaucgucaacauccaTT 560 MP
AL-DP-8144 ggauguugacgauggucauTT 561 augaccaucgucaacauccTT 562 MP
AL-DP-8145 gauguugacgauggucauuTT 563 aaugaccaucgucaacaucTT 564 MP
AL-DP-8146 auguugacgauggucauuuTT 565 aaaugaccaucgucaacauTT 566 MP
AL-DP-8147 uguugacgauggucauuuuTT 567 aaaaugaccaucgucaacaTT 568 MP
AL-DP-8148 guugacgauggucauuuugTT 569 caaaaugaccaucgucaacTT 570 MP
AL-DP-8149 uugacgauggucauuuuguTT 571 acaaaaugaccaucgucaaTT 572 MP
AL-DP-8152 acgauggucauuuugucaaTT 573 uugacaaaaugaccaucguTT 574 MP
AL-DP-8153 cgauggucauuuugucaacTT 575 guugacaaaaugaccaucgTT 576 MP
AL-DP-8154 gauggucauuuugucaacaTT 577 uguugacaaaaugaccaucTT 578 MP
AL-DP-8155 auggucauuuugucaacauTT 579 auguugacaaaaugaccauTT 580 MP
AL-DP-8156 uggucauuuugucaacauaTT 581 uauguugacaaaaugaccaTT 582 MP
AL-DP-8157 ggucauuuugucaacauagTT 583 cuauguugacaaaaugaccTT 584 MP
AL-DP-8158 gucauuuugucaacauagaTT 585 ucuauguugacaaaaugacTT 586 MP
AL-DP-8159 ucaaggcaccaaacgaucuTT 587 agaucguuuggugccuugaTT 588 NP
AL-DP-8160 caaggcaccaaacgaucuuTT 589 aagaucguuuggugccuugTT 590 NP
AL-DP-8161 aaggcaccaaacgaucuuaTT 591 uaagaucguuuggugccuuTT 592 NP
AL-DP-8162 aacagcauaacaauagagaTT 593 ucucuauuguuaugcuguuTT 594 NP
AL-DP-8163 acagcauaacaauagagagTT 595 cucucuauuguuaugcuguTT 596 NP
AL-DP-8164 cagcauaacaauagagagaTT 597 ucucucuauuguuaugcugTT 598 NP
AL-DP-8165 agcauaacaauagagagaaTT 599 uucucucuauuguuaugcuTT 600 NP
AL-DP-8166 gcauaacaauagagagaauTT 601 auucucucuauuguuaugcTT 602 NP
AL-DP-8167 cauaacaauagagagaaugTT 603 cauucucucuauuguuaugTT 604 NP
AL-DP-8168 auugcauaugagagaauguTT 605 acauucucucauaugcaauTT 606 NP
AL-DP-8169 uugcauaugagagaaugugTT 607 cacauucucucauaugcaaTT 608 NP
AL-DP-8170 gcauaugagagaaugugcaTT 609 ugcacauucucucauaugcTT 610 NP
AL-DP-8171 cauaugagagaaugugcaaTT 611 uugcacauucucucauaugTT 612 NP
AL-DP-8177 gggagucuucgagcucucgTT 613 cgagagcucgaagacucccTT 614 NP
AL-DP-8178 ggagucuucgagcucucggTT 615 ccgagagcucgaagacuccTT 616 NP
AL-DP-8179 gagucuucgagcucucggaTT 617 uccgagagcucgaagacucTT 618 NP
AL-DP-8180 agucuucgagcucucggacTT 619 guccgagagcucgaagacuTT 620 NP
AL-DP-8181 gucuucgagcucucggacgTT 621 cguccgagagcucgaagacTT 622 NP
AL-DP-8182 ucuucgagcucucggacgaTT 623 ucguccgagagcucgaagaTT 624 NP
AL-DP-8183 cuucgagcucucggacgaaTT 625 uucguccgagagcucgaagTT 626 NP
AL-DP-8184 uucgagcucucggacgaaaTT 627 uuucguccgagagcucgaaTT 628 NP
AL-DP-8185 cgagcucucggacgaaaagTT 629 cuuuucguccgagagcucgTT 630 NP
AL-DP-8186 gagcucucggacgaaaaggTT 631 ccuuuucguccgagagcucTT 632 NP
AL-DP-8187 agcucucggacgaaaaggcTT 633 gccuuuucguccgagagcuTT 634 NP
AL-DP-8188 gcucucggacgaaaaggcaTT 635 ugccuuuucguccgagagcTT 636 NP
AL-DP-8189 cucucggacgaaaaggcaaTT 637 uugccuuuucguccgagagTT 638 NP
AL-DP-8190 ucucggacgaaaaggcaacTT 639 guugccuuuucguccgagaTT 640 NP
AL-DP-8191 cucggacgaaaaggcaacgTT 641 cguugccuuuucguccgagTT 642 NP
AL-DP-8192 ucggacgaaaaggcaacgaTT 643 ucguugccuuuucguccgaTT 644 NP
AL-DP-8193 cggacgaaaaggcaacgaaTT 645 uucguugccuuuucguccgTT 646 NP
AL-DP-8194 ggacgaaaaggcaacgaacTT 647 guucguugccuuuucguccTT 648 NP
AL-DP-8195 gacgaaaaggcaacgaaccTT 649 gguucguugccuuuucgucTT 650 NP
AL-DP-8196 acgaaaaggcaacgaacccTT 651 ggguucguugccuuuucguTT 652 NP
AL-DP-8197 cgaaaaggcaacgaacccgTT 653 cggguucguugccuuuucgTT 654 NP
AL-DP-8198 aaaaggcaacgaacccgauTT 655 aucggguucguugccuuuuTT 656 NP
AL-DP-8199 aaaggcaacgaacccgaucTT 657 gaucggguucguugccuuuTT 658 NP
AL-DP-8200 aaggcaacgaacccgaucgTT 659 cgaucggguucguugccuuTT 660 NP
AL-DP-8201 acgaacccgaucgugccuuTT 661 aaggcacgaucggguucguTT 662 NP
AL-DP-8202 cgaacccgaucgugccuucTT 663 gaaggcacgaucggguucgTT 664 NP
AL-DP-8203 gaacccgaucgugccuuccTT 665 ggaaggcacgaucggguucTT 666 NP
AL-DP-8204 aacccgaucgugccuuccuTT 667 aggaaggcacgaucggguuTT 668 NP
AL-DP-8205 acccgaucgugccuuccuuTT 669 aaggaaggcacgaucggguTT 670 NP
AL-DP-8221 auggaugucaauccgacuuTT 671 aagucggauugacauccauTT 672 PB1
AL-DP-8222 uggaugucaauccgacuuuTT 673 aaagucggauugacauccaTT 674 PB1
AL-DP-8223 gaugucaauccgacuuuacTT 675 guaaagucggauugacaucTT 676 PB1
AL-DP-8224 augucaauccgacuuuacuTT 677 aguaaagucggauugacauTT 678 PB1
AL-DP-8225 ugucaauccgacuuuacuuTT 679 aaguaaagucggauugacaTT 680 PB1
AL-DP-8226 ccauacagccauggaacagTT 681 cuguuccauggcuguauggTT 682 PB1
AL-DP-8227 cauacagccauggaacaggTT 683 ccuguuccauggcuguaugTT 684 PB1
AL-DP-8228 acaggauacaccauggacaTT 685 uguccaugguguauccuguTT 686 PB1
AL-DP-8229 caggauacaccauggacacTT 687 guguccaugguguauccugTT 688 PB1
AL-DP-8230 uuggaagcaauggcuuuccTT 689 ggaaagccauugcuuccaaTT 690 PB1
AL-DP-8231 ggaagcaauggcuuuccuuTT 691 aaggaaagccauugcuuccTT 692 PB1
AL-DP-8232 agcaauggcuuuccuugaaTT 693 uucaaggaaagccauugcuTT 694 PB1
AL-DP-8233 augaugacuaacucacaagTT 695 cuugugaguuagucaucauTT 696 PB1
AL-DP-8234 ugaugacuaacucacaagaTT 697 ucuugugaguuagucaucaTT 698 PB1
AL-DP-8235 accaaauggaaugagaaucTT 699 gauucucauuccauuugguTT 700 PB1
AL-DP-8236 ccaaauggaaugagaaucaTT 701 ugauucucauuccauuuggTT 702 PB1
AL-DP-8237 ggaaugaugaugggcauguTT 703 acaugcccaucaucauuccTT 704 PB1
AL-DP-8238 gaaugaugaugggcauguuTT 705 aacaugcccaucaucauucTT 706 PB1
AL-DP-8239 cuccaauccucugaugauuTT 707 aaucaucagaggauuggagTT 708 PB1
AL-DP-8240 uccaauccucugaugauuuTT 709 aaaucaucagaggauuggaTT 710 PB1
AL-DP-8241 aucaugagggaauacaagcTT 711 gcuuguauucccucaugauTT 712 PB1
AL-DP-8242 uuugugcgacaaugcuucaTT 713 ugaagcauugucgcacaaaTT 714 PA
AL-DP-8243 uaugggauuccuuucgucaTT 715 ugacgaaaggaaucccauaTT 716 PA
AL-DP-8244 uccgagagaggcgaagagaTT 717 ucucuucgccucucucggaTT 718 PA
AL-DP-8245 ccgagagaggcgaagagacTT 719 gucucuucgccucucucggTT 720 PA
AL-DP-8247 gagagaggcgaagagacaaTT 721 uugucucuucgccucucucTT 722 PA
AL-DP-8248 agagaggcgaagagacaauTT 723 auugucucuucgccucucuTT 724 PA
AL-DP-8249 gagaggcgaagagacaauuTT 725 aauugucucuucgccucucTT 726 PA
AL-DP-8250 agaggcgaagagacaauugTT 727 caauugucucuucgccucuTT 728 PA
AL-DP-8251 gaggcgaagagacaauugaTT 729 ucaauugucucuucgccucTT 730 PA
AL-DP-8252 aggcgaagagacaauugaaTT 731 uucaauugucucuucgccuTT 732 PA
AL-DP-8253 ggcgaagagacaauugaagTT 733 cuucaauugucucuucgccTT 734 PA
AL-DP-8254 gcgaagagacaauugaagaTT 735 ucuucaauugucucuucgcTT 736 PA
AL-DP-8255 cgaagagacaauugaagaaTT 737 uucuucaauugucucuucgTT 738 PA
AL-DP-8257 uucuccagccuugaaaacuTT 739 aguuuucaaggcuggagaaTT 740 PA
AL-DP-8258 ucuccagccuugaaaacuuTT 741 aaguuuucaaggcuggagaTT 742 PA
AL-DP-8259 cuccagccuugaaaacuuuTT 743 aaaguuuucaaggcuggagTT 744 PA
AL-DP-8260 uccagccuugaaaacuuuaTT 745 uaaaguuuucaaggcuggaTT 746 PA
AL-DP-8261 ccagccuugaaaacuuuagTT 747 cuaaaguuuucaaggcuggTT 748 PA
AL-DP-8262 cagccuugaaaacuuuagaTT 749 ucuaaaguuuucaaggcugTT 750
PA
AL-DP-8263 agccuugaaaacuuuagagTT 751 cucuaaaguuuucaaggcuTT 752 PA
AL-DP-8264 gccuugaaaacuuuagagcTT 753 gcucuaaaguuuucaaggcTT 754 PA
AL-DP-8265 ccuugaaaacuuuagagccTT 755 ggcucuaaaguuuucaaggTT 756 PA
AL-DP-8266 cuugaaaacuuuagagccuTT 757 aggcucuaaaguuuucaagTT 758 PA
AL-DP-8267 uugaaaacuuuagagccuaTT 759 uaggcucuaaaguuuucaaTT 760 PA
AL-DP-8268 ugaaaacuuuagagccuauTT 761 auaggcucuaaaguuuucaTT 762 PA
AL-DP-8269 gaaaacuuuagagccuaugTT 763 cauaggcucuaaaguuuucTT 764 PA
AL-DP-8270 aaaacuuuagagccuauguTT 765 acauaggcucuaaaguuuuTT 766 PA
AL-DP-8271 aaacuuuagagccuaugugTT 767 cacauaggcucuaaaguuuTT 768 PA
AL-DP-8272 aacuuuagagccuauguggTT 769 ccacauaggcucuaaaguuTT 770 PA
AL-DP-8273 acuuuagagccuauguggaTT 771 uccacauaggcucuaaaguTT 772 PA
AL-DP-8274 cuuuagagccuauguggauTT 773 auccacauaggcucuaaagTT 774 PA
AL-DP-8281 aaccgaacggcugcauugaTT 775 ucaaugcagccguucgguuTT 776 PA
AL-DP-8282 accgaacggcugcauugagTT 777 cucaaugcagccguucgguTT 778 PA
AL-DP-8283 ccgaacggcugcauugaggTT 779 ccucaaugcagccguucggTT 780 PA
AL-DP-8284 cgaacggcugcauugagggTT 781 cccucaaugcagccguucgTT 782 PA
AL-DP-8285 gaacggcugcauugagggcTT 783 gcccucaaugcagccguucTT 784 PA
AL-DP-8286 aacggcugcauugagggcaTT 785 ugcccucaaugcagccguuTT 786 PA
AL-DP-8287 acggcugcauugagggcaaTT 787 uugcccucaaugcagccguTT 788 PA
AL-DP-8288 cggcugcauugagggcaagTT 789 cuugcccucaaugcagccgTT 790 PA
AL-DP-8289 ggcugcauugagggcaagcTT 791 gcuugcccucaaugcagccTT 792 PA
AL-DP-8290 gcugcauugagggcaagcuTT 793 agcuugcccucaaugcagcTT 794 PA
AL-DP-8291 cugcauugagggcaagcuuTT 795 aagcuugcccucaaugcagTT 796 PA
AL-DP-8292 ugcauugagggcaagcuuuTT 797 aaagcuugcccucaaugcaTT 798 PA
AL-DP-8293 gcauugagggcaagcuuucTT 799 gaaagcuugcccucaaugcTT 800 PA
AL-DP-8294 cauugagggcaagcuuucuTT 801 agaaagcuugcccucaaugTT 802 PA
AL-DP-8295 auugagggcaagcuuucucTT 803 gagaaagcuugcccucaauTT 804 PA
AL-DP-8296 uugagggcaagcuuucucaTT 805 ugagaaagcuugcccucaaTT 806 PA
AL-DP-8297 ugagggcaagcuuucucaaTT 807 uugagaaagcuugcccucaTT 808 PA
AL-DP-8298 gagggcaagcuuucucaaaTT 809 uuugagaaagcuugcccucTT 810 PA
AL-DP-8299 agggcaagcuuucucaaauTT 811 auuugagaaagcuugcccuTT 812 PA
AL-DP-8300 gggcaagcuuucucaaaugTT 813 cauuugagaaagcuugcccTT 814 PA
AL-DP-8301 ggcaagcuuucucaaauguTT 815 acauuugagaaagcuugccTT 816 PA
AL-DP-8302 gcaagcuuucucaaaugucTT 817 gacauuugagaaagcuugcTT 818 PA
AL-DP-8303 augauaagcaaaugcagaaTT 819 uucugcauuugcuuaucauTT 820 PA
AL-DP-8304 ugauaagcaaaugcagaacTT 821 guucugcauuugcuuaucaTT 822 PA
AL-DP-8305 cuuagggacaaccuggaacTT 823 guuccagguugucccuaagTT 824 PA
AL-DP-8306 uuagggacaaccuggaaccTT 825 gguuccagguugucccuaaTT 826 PA
AL-DP-8307 uagggacaaccuggaaccuTT 827 agguuccagguugucccuaTT 828 PA
AL-DP-8308 agggacaaccuggaaccugTT 829 cagguuccagguugucccuTT 830 PA
AL-DP-8309 gggacaaccuggaaccuggTT 831 ccagguuccagguugucccTT 832 PA
AL-DP-8315 agcaauugaggagugccugTT 833 caggcacuccucaauugcuTT 834 PA
AL-DP-8316 gcaauugaggagugccugaTT 835 ucaggcacuccucaauugcTT 836 PA
AL-DP-8317 caauugaggagugccugauTT 837 aucaggcacuccucaauugTT 838 PA
AL-DP-8318 aauugaggagugccugauuTT 839 aaucaggcacuccucaauuTT 840 PA
AL-DP-8319 auugaggagugccugauuaTT 841 uaaucaggcacuccucaauTT 842 PA
AL-DP-8320 uugaggagugccugauuaaTT 843 uuaaucaggcacuccucaaTT 844 PA
AL-DP-8328 uugguucaacuccuuccucTT 845 gaggaaggaguugaaccaaTT 846 PA
AL-DP-8329 gaaugaaauggaugauggcTT 847 gccaucauccauuucauucTT 848 PB2
AL-DP-8330 ugguaaugaaacggaaacgTT 849 cguuuccguuucauuaccaTT 850 PB2
AL-DP-8331 augaaacggaaacgggacuTT 851 agucccguuuccguuucauTT 852 PB2
AL-DP-8332 ugaaacggaaacgggacucTT 853 gagucccguuuccguuucaTT 854 PB2
AL-DP-8333 gaaacggaaacgggacucuTT 855 agagucccguuuccguuucTT 856 PB2
AL-DP-8334 aaacggaaacgggacucuaTT 857 uagagucccguuuccguuuTT 858 PB2
AL-DP-8335 aacggaaacgggacucuagTT 859 cuagagucccguuuccguuTT 860 PB2
AL-DP-8336 acggaaacgggacucuagcTT 861 gcuagagucccguuuccguTT 862 PB2
AL-DP-8337 cggaaacgggacucuagcaTT 863 ugcuagagucccguuuccgTT 864 PB2
AL-DP-8338 ggaaacgggacucuagcauTT 865 augcuagagucccguuuccTT 866 PB2
.sup.1in in vitro plaque forming assay in MCDK cells as described
in Example 3.1; .sup.2in in vitro ELISA assay in MCDK cells as
described in Example 3.2: .sup.3in in vitro ELISA assay in MCDK
cells as described in Example 3.2; .sup.4in in vitro ELISA assay in
MCDK cells as described in Example 3.2; negative values indicate
that target gene expression was enhanced in treated cells compared
to controls
[0053] TABLE-US-00006 TABLE 1F Exemplary iRNA agents for targeting
influenza virus,and having 100% target coverage (criterium 1, see
example 1) and 100% target efficiency (criterium 2, see example 1),
but allowing for up to 3 universal bases in the non-seed region of
the iRNA agent; x stands for the position of a universal base SEQ
SEQ Target Duplex Sense strand sequence ID Antisense strand ID
influenza identifier (5'-3') NO: sequence (5'-3') NO: gene
AL-DP-8368 ucxgcxgaxaugagcauugTT 867 caaugcucauxucxgcxgaTT 868 PB1
AL-DP-8369 cxgcxgaxaugagcauuggTT 869 ccaaugcucauxucxgcxgTT 870 PB1
AL-DP-8370 xaagaucugxuccaccauuTT 871 aaugguggaxcagaucuuxTT 872 PB1
AL-DP-8371 aagaucugxuccaccauugTT 873 caaugguggaxcagaucuuTT 874 PB1
AL-DP-8372 agaucugxuccaccauugxTT 875 xcaaugguggaxcagaucuTT 876 PB1
AL-DP-8373 xugaauuuxxcuuguccuuTT 877 aaggacaagxxaaauucaxTT 878 PB1
AL-DP-8374 ugaauuuxxcuuguccuucTT 879 gaaggacaagxxaaauucaTT 880 PB1
AL-DP-8375 gaauuuxxcuuguccuucxTT 881 xgaaggacaagxxaaauucTT 882 PB1
AL-DP-8376 uuguccuucxugaaaaaauTT 883 auuuuuucaxgaaggacaaTT 884 PB1
AL-DP-8377 uguccuucxugaaaaaaugTT 885 cauuuuuucaxgaaggacaTT 886 PB1
AL-DP-8378 guccuucxugaaaaaaugcTT 887 gcauuuuuucaxgaaggacTT 888 PB1
AL-DP-8379 uccuucxugaaaaaaugcxTT 889 xgcauuuuuucaxgaaggaTT 890 PB1
AL-DP-8380 aaxggxugxauugagggcaTT 891 ugcccucaauxcaxccxuuTT 892 PA
AL-DP-8381 axggxugxauugagggcaaTT 893 uugcccucaauxcaxccxuTT 894 PA
AL-DP-8382 xggxugxauugagggcaagTT 895 cuugcccucaauxcaxccxTT 896 PA
AL-DP-8383 ggxugxauugagggcaagcTT 897 gcuugcccucaauxcaxccTT 898 PA
AL-DP-8384 gxugxauugagggcaagcuTT 899 agcuugcccucaauxcaxcTT 900 PA
AL-DP-8385 xugxauugagggcaagcuwTT 901 wagcuugcccucaauxcaxTT 902 PA
AL-DP-8386 aaxgcuacuxuuugcuaucTT 903 gauagcaaaxaguagcxuuTT 904 PA
AL-DP-8387 axgcuacuxuuugcuauccTT 905 ggauagcaaaxaguagcxuTT 906 PA
AL-DP-8388 xgcuacuxuuugcuauccaTT 907 uggauagcaaaxaguagcxTT 908 PA
AL-DP-8389 gcuacuxuuugcuauccauTT 909 auggauagcaaaxaguagcTT 910 PA
AL-DP-8390 cuacuxuuugcuauccauaTT 911 uauggauagcaaaxaguagTT 912 PA
AL-DP-8391 uacuxuuugcuauccauacTT 913 guauggauagcaaaxaguaTT 914 PA
AL-DP-8392 acuxuuugcuauccauacuTT 915 aguauggauagcaaaxaguTT 916 PA
AL-DP-8393 cuxuuugcuauccauacugTT 917 caguauggauagcaaaxagTT 918 PA
AL-DP-8394 uxuuugcuauccauacuguTT 919 acaguauggauagcaaaxaTT 920 PA
AL-DP-8395 xuuugcuauccauacugucTT 921 gacaguauggauagcaaaxTT 922 PA
AL-DP-8396 uuugcuauccauacugucxTT 923 xgacaguauggauagcaaaTT 924 PA
AL-DP-8397 ucggccxxcxaaagcagguTT 925 accugcuuuxgxxggccgaTT 926 PB2
AL-DP-8398 cggccxxcxaaagcaggucTT 927 gaccugcuuuxgxxggccgTT 928 PB2
AL-DP-8399 ggccxxcxaaagcaggucaTT 929 ugaccugcuuuxgxxggccTT 930 PB2
AL-DP-8400 gccxxcxaaagcaggucaaTT 931 uugaccugcuuuxgxxggcTT 932 PB2
AL-DP-8401 gacagxcagxcagcgaccaTT 933 uggucgcugxcugxcugucTT 934 PB2
AL-DP-8402 acagxcagxcagcgaccaxTT 935 xuggucgcugxcugxcuguTT 936 PB2
AL-DP-8403 xuxuhgaauxguuuaaaaaTT 937 uuuuuaaacxauuchaxaxTT 938 PB2
AL-DP-8404 uguxgaauxguuuaaaaacTT 939 guuuuuaaacxauucxacaTT 940 PB2
AL-DP-8405 guxgaauxguuuaaaaacsTT 941 sguuuuuaaacxauucxacTT 942 PB2
AL-DP-8406 uguuucuxxxauauggcgcTT 943 gcgccauauxxxagaaacaTT 944 PB2
AL-DP-8407 guuucuxxxauauggcgcaTT 945 ugcgccauauxxxagaaacTT 946 PB2
AL-DP-8408 uuucuxxxauauggcgcauTT 947 augcgccauauxxxagaaaTT 948 PB2
AL-DP-8409 uucuxxxauauggcgcauaTT 949 uaugcgccauauxxxagaaTT 950 PB2
AL-DP-8410 ucuxxxauauggcgcauacTT 951 guaugcgccauauxxxagaTT 952 PB2
AL-DP-8411 cuxxxauauggcgcauacuTT 953 aguaugcgccauauxxxagTT 954 PB2
AL-DP-8412 uxxxauauggcgcauacucTT 955 gaguaugcgccauauxxxaTT 956 PB2
AL-DP-8413 xxxuauggcgcauacucgTT 957 cgaguaugcgccauaxxxTT 958 PB2
AL-DP-8414 xxauauggcgcauacucggTT 959 ccgaguaugcgccauauxxTT 960 PB2
AL-DP-8415 xauauggcgcauacucgggTT 961 cccgaguaugcgccauauxTT 962 PB2
AL-DP-8416 auauggcgcauacucgggcTT 963 gcccgaguaugcgccauauTT 964 PB2
AL-DP-8417 uauggcgcauacucgggcaTT 965 ugcccgaguaugcgccauaTT 966 PB2
AL-DP-8418 auggcgcauacucgggcauTT 967 augcccgaguaugcgccauTT 968 PB2
AL-DP-8419 uggcgcauacucgggcaugTT 969 caugcccgaguaugcgccaTT 970 PB2
AL-DP-8420 ggcgcauacucgggcauguTT 971 acaugcccgaguaugcgccTT 972 PB2
AL-DP-8421 xxggcccccxcaaagccgaTT 973 ucggcuuugxgggggccxxTT 974 MP
AL-DP-8422 xggcccccxcaaagccgaxTT 975 xucggcuuugxgggggccxTT 976 MP
AL-DP-8423 xuuxacgcuxaccgugcccTT 977 gggcacgguxagcguxaaxTT 978 MP
AL-DP-8424 uuxacgcuxaccgugcccaTT 979 ugggcacgguxagcguxaaTT 980 MP
AL-DP-8425 uxacgcuxaccgugcccagTT 981 cugggcacgguxagcguxaTT 982 MP
AL-DP-8426 xacgcuxaccgugcccagxTT 983 xcugggcacgguxagcguxTT 984 MP
.sup.1in in vitro plaque forming assay in MCDK cells as described
in Example 3.1
[0054] TABLE-US-00007 TABLE 1G Exemplary iRNA agents for targeting
influenza virus,and having 80% target coverage (criterium 1, see
example 1) and 80% target efficiency (criterium 2, see example 1),
but allowing for 1 universal base in the non-seed region of the
iRNA agent; x stands for the position of a universal base SEQ SEQ
Target Duplex Sense strand sequence ID Antisense strand ID
influenza identifier (5'-3') NO: sequence (5'-3') NO: gene
AL-DP-8427 acguacguucucucuaucxTT 985 xgauagagagaacguacguTT 986 MP
AL-DP-8428 cucucuaucxucccgucagTT 987 cugacgggaxgauagagagTT 988 MP
AL-DP-8429 ucucuaucxucccgucaggTT 989 ccugacgggaxgauagagaTT 990 MP
AL-DP-8430 cucuaucxucccgucaggcTT 991 gccugacgggaxgauagagTT 992 MP
AL-DP-8431 ucuaucxucccgucaggccTT 993 ggccugacgggaxgauagaTT 994 MP
AL-DP-8432 cuaucxucccgucaggcccTT 995 gggccugacgggaxgauagTT 996 MP
AL-DP-8433 uaucxucccgucaggccccTT 997 ggggccugacgggaxgauaTT 998 MP
AL-DP-8434 aucxucccgucaggcccccTT 999 gggggccugacgggaxgauTT 1000 MP
AL-DP-8435 ucxucccgucaggcccccuTT 1001 agggggccugacgggaxgaTT 1002 MP
AL-DP-8436 cxucccgucaggcccccucTT 1003 gagggggccugacgggaxgTT 1004 MP
AL-DP-8437 xucccgucaggcccccucaTT 1005 ugagggggccugacgggaxTT 1006 MP
AL-DP-8438 aagccgagaucgcgcagaxTT 1007 xucugcgcgaucucggcuuTT 1008 MP
AL-DP-8439 gaucuxgaggcucucauggTT 1009 ccaugagagccucxagaucTT 1010 MP
AL-DP-8440 aucuxgaggcucucauggaTT 1011 uccaugagagccucxagauTT 1012 MP
AL-DP-8441 ucucauggaxuggcuaaagTT 1013 cuuuagccaxuccaugagaTT 1014 MP
AL-DP-8442 cucauggaxuggcuaaagaTT 1015 ucuuuagccaxuccaugagTT 1016 MP
AL-DP-8443 ucauggaxuggcuaaagacTT 1017 gucuuuagccaxuccaugaTT 1018 MP
AL-DP-8444 cauggaxuggcuaaagacaTT 1019 ugucuuuagccaxuccaugTT 1020 MP
AL-DP-8445 auggaxuggcuaaagacaaTT 1021 uugucuuuagccaxuccauTT 1022 MP
AL-DP-8446 uggaxuggcuaaagacaagTT 1023 cuugucuuuagccaxuccaTT 1024 MP
AL-DP-8447 ggaxuggcuaaagacaagaTT 1025 ucuugucuuuagccaxuccTT 1026 MP
AL-DP-8448 gaxuggcuaaagacaagacTT 1027 gucuugucuuuagccaxucTT 1028 MP
AL-DP-8449 axuggcuaaagacaagaccTT 1029 ggucuugucuuuagccaxuTT 1030 MP
AL-DP-8450 xuggcuaaagacaagaccaTT 1031 uggucuugucuuuagccaxTT 1032 MP
AL-DP-8451 ccugucaccucugacuaaxTT 1033 xuuagucagaggugacaggTT 1034 MP
AL-DP-8452 uuuguxuucacgcucaccgTT 1035 cggugagcgugaaxacaaaTT 1036 MP
AL-DP-8453 uuguxuucacgcucaccguTT 1037 acggugagcgugaaxacaaTT 1038 MP
AL-DP-8454 uguxuucacgcucaccgugTT 1039 cacggugagcgugaaxacaTT 1040 MP
AL-DP-8455 guxuucacgcucaccgugcTT 1041 gcacggugagcgugaaxacTT 1042 MP
AL-DP-8456 uxuucacgcucaccgugccTT 1043 ggcacggugagcgugaaxaTT 1044 MP
AL-DP-8457 xuucacgcucaccgugcccTT 1045 gggcacggugagcgugaaxTT 1046 MP
AL-DP-8458 aggacugcagcguagacgxTT 1047 xcgucuacgcugcaguccuTT 1048 MP
AL-DP-8459 ugxaugggucucauauacaTT 1049 uguauaugagacccauxcaTT 1050 MP
AL-DP-8460 gxaugggucucauauacaaTT 1051 uuguauaugagacccauxcTT 1052 MP
AL-DP-8461 xaugggucucauauacaacTT 1053 guuguauaugagacccauxTT 1054 MP
AL-DP-8462 augggucucauauacaacxTT 1055 xguuguauaugagacccauTT 1056 MP
AL-DP-8463 ugugccacxugugagcagaTT 1057 ucugcucacaxguggcacaTT 1058 MP
AL-DP-8464 gugccacxugugagcagauTT 1059 aucugcucacaxguggcacTT 1060 MP
AL-DP-8465 gccacxugugagcagauugTT 1061 caaucugcucacaxguggcTT 1062 MP
AL-DP-8466 ccacxugugagcagauugcTT 1063 gcaaucugcucacaxguggTT 1064 MP
AL-DP-8467 cuaggcaxauggugcaggcTT 1065 gccugcaccauxugccuagTT 1066 MP
AL-DP-8468 augagxacaauugggacucTT 1067 gagucccaauuguxcucauTT 1068 MP
AL-DP-8469 ugagxacaauugggacucaTT 1069 ugagucccaauuguxcucaTT 1070 MP
AL-DP-8470 uuugcaggcxuaccagaaaTT 1071 uuucugguaxgccugcaaaTT 1072 MP
AL-DP-8471 uugcaggcxuaccagaaacTT 1073 guuucugguaxgccugcaaTT 1074 MP
AL-DP-8472 ugcaggcxuaccagaaacgTT 1075 cguuucugguaxgccugcaTT 1076 MP
AL-DP-8473 augcagcgxuucaagugauTT 1077 aucacuugaaxcgcugcauTT 1078 MP
AL-DP-8474 ugcagcgxuucaagugaucTT 1079 gaucacuugaaxcgcugcaTT 1080 MP
AL-DP-8475 gcagcgxuucaagugauccTT 1081 ggaucacuugaaxcgcugcTT 1082 MP
AL-DP-8476 cagcgxuucaagugauccuTT 1083 aggaucacuugaaxcgcugTT 1084 MP
AL-DP-8477 agcgxuucaagugauccucTT 1085 gaggaucacuugaaxcgcuTT 1086 MP
AL-DP-8478 gcgxuucaagugauccucuTT 1087 agaggaucacuugaaxcgcTT 1088 MP
AL-DP-8479 cauugggauxuugcacuugTT 1089 caagugcaaxaucccaaugTT 1090 MP
AL-DP-8480 auugggauxuugcacuugaTT 1091 ucaagugcaaxaucccaauTT 1092 MP
AL-DP-8481 uugggauxuugcacuugauTT 1093 aucaagugcaaxaucccaaTT 1094 MP
AL-DP-8482 ugggauxuugcacuugauaTT 1095 uaucaagugcaaxaucccaTT 1096 MP
AL-DP-8483 gggauxuugcacuugauauTT 1097 auaucaagugcaaxaucccTT 1098 MP
AL-DP-8484 ggauxuugcacuugauauuTT 1099 aauaucaagugcaaxauccTT 1100 MP
AL-DP-8485 gauxuugcacuugauauugTT 1101 caauaucaagugcaaxaucTT 1102 MP
AL-DP-8486 auxuugcacuugauauuguTT 1103 acaauaucaagugcaaxauTT 1104 MP
AL-DP-8487 uxuugcacuugauauugugTT 1105 cacaauaucaagugcaaxaTT 1106 MP
AL-DP-8488 xuugcacuugauauuguggTT 1107 ccacaauaucaagugcaaxTT 1108 MP
AL-DP-8489 aaaugcauuuaucgucgcxTT 1109 xgcgacgauaaaugcauuuTT 1110 MP
AL-DP-8490 uaucgucgcxuuaaauacgTT 1111 cguauuuaaxgcgacgauaTT 1112 MP
AL-DP-8491 aucgucgcxuuaaauacggTT 1113 ccguauuuaaxgcgacgauTT 1114 MP
AL-DP-8492 ucgucgcxuuaaauacgguTT 1115 accguauuuaaxgcgacgaTT 1116 MP
AL-DP-8493 cgucgcxuuaaauacgguuTT 1117 aaccguauuuaaxgcgacgTT 1118 MP
AL-DP-8494 gucgcxuuaaauacgguuuTT 1119 aaaccguauuuaaxgcgacTT 1120 MP
AL-DP-8495 ucgcxuuaaauacgguuugTT 1121 caaaccguauuuaaxgcgaTT 1122 MP
AL-DP-8496 cgcxuuaaauacgguuugaTT 1123 ucaaaccguauuuaaxgcgTT 1124 MP
AL-DP-8497 gcxuuaaauacgguuugaaTT 1125 uucaaaccguauuuaaxgcTT 1126 MP
AL-DP-8498 cxuuaaauacgguuugaaaTT 1127 uuucaaaccguauuuaaxgTT 1128 MP
AL-DP-8499 xuuaaauacgguuugaaaaTT 1129 uuuucaaaccguauuuaaxTT 1130 MP
AL-DP-8500 gguuugaaaagagggccuxTT 1131 xaggcccucuuuucaaaccTT 1132 MP
AL-DP-8501 xuugaaaagagggccuucuTT 1133 agaaggcccucuuuucaaxTT 1134 MP
AL-DP-8502 aaaaxagggccuucuacggTT 1135 ccguagaaggcccuxuuuuTT 1136 MP
AL-DP-8503 aaagagggccuucuacggxTT 1137 xccguagaaggcccucuuuTT 1138 MP
AL-DP-8504 guxccugagucuaugagggTT 1139 cccucauagacucaggxacTT 1140 MP
AL-DP-8505 uxccugagucuaugagggaTT 1141 ucccucauagacucaggxaTT 1142 MP
AL-DP-8506 xccugagucuaugagggaaTT 1143 uucccucauagacucaggxTT 1144 MP
AL-DP-8507 ugagucuaugagggaagaxTT 1145 xucuucccucauagacucaTT 1146 MP
AL-DP-8508 cagaxugcuguggauguugTT 1147 caacauccacagcaxucugTT 1148 MP
AL-DP-8509 agaxugcuguggauguugaTT 1149 ucaacauccacagcaxucuTT 1150 MP
AL-DP-8510 gaxugcuguggauguugacTT 1151 gucaacauccacagcaxucTT 1152 MP
AL-DP-8511 axugcuguggauguugacgTT 1153 cgucaacauccacagcaxuTT 1154 MP
AL-DP-8512 xugcuguggauguugacgaTT 1155 ucgucaacauccacagcaxTT 1156 MP
AL-DP-8513 xucaaggcaccaaacgaucTT 1157 gaucguuuggugccuugaxTT 1158 NP
AL-DP-8514 aggcaccaaacgaucuuaxTT 1159 xuaagaucguuuggugccuTT 1160 NP
AL-DP-8515 uaugaxcagauggaaacugTT 1161 caguuuccaucugxucauaTT 1162 NP
AL-DP-8516 augaxcagauggaaacuggTT 1163 ccaguuuccaucugxucauTT 1164 NP
AL-DP-8517 ugaxcagauggaaacugguTT 1165 accaguuuccaucugxucaTT 1166 NP
AL-DP-8518 gaxcagauggaaacuggugTT 1167 caccaguuuccaucugxucTT 1168 NP
AL-DP-8519 axcagauggaaacugguggTT 1169 ccaccaguuuccaucugxuTT 1170 NP
AL-DP-8520 aacugguggxgaacgccagTT 1171 cuggcguucxccaccaguuTT 1172 NP
AL-DP-8521 acugguggxgaacgccagaTT 1173 ucuggcguucxccaccaguTT 1174 NP
AL-DP-8522 cugguggxgaacgccagaaTT 1175 uucuggcguucxccaccagTT 1176 NP
AL-DP-8523 ugguggxgaacgccagaauTT 1177 auucuggcguucxccaccaTT 1178 NP
AL-DP-8524 gguggxgaacgccagaaugTT 1179 cauucuggcguucxccaccTT 1180 NP
AL-DP-8525 guggxgaacgccagaaugcTT 1181 gcauucuggcguucxccacTT 1182 NP
AL-DP-8526 ccagaaugcxacugagaucTT 1183 gaucucaguxgcauucuggTT 1184 NP
AL-DP-8527 cagaaugcxacugagaucaTT 1185 ugaucucaguxgcauucugTT 1186 NP
AL-DP-8528 agaaugcxacugagaucagTT 1187 cugaucucaguxgcauucuTT 1188 NP
AL-DP-8529 gaugugcacxgaacucaaaTT 1189 uuugaguucxgugcacaucTT 1190 NP
AL-DP-8530 augugcacxgaacucaaacTT 1191 guuugaguucxgugcacauTT 1192 NP
AL-DP-8531 ugugcacxgaacucaaacuTT 1193 aguuugaguucxgugcacaTT 1194 NP
AL-DP-8532 gugcacxgaacucaaacucTT 1195 gaguuugaguucxgugcacTT 1196 NP
AL-DP-8533 ugcacxgaacucaaacucaTT 1197 ugaguuugaguucxgugcaTT 1198 NP
AL-DP-8534 gcacxgaacucaaacucagTT 1199 cugaguuugaguucxgugcTT 1200 NP
AL-DP-8535 caxaacagcauaacaauagTT 1201 cuauuguuaugcuguuxugTT 1202 NP
AL-DP-8536 axaacagcauaacaauagaTT 1203 ucuauuguuaugcuguuxuTT 1204 NP
AL-DP-8537 xaacagcauaacaauagagTT 1205 cucuauuguuaugcuguuxTT 1206 NP
AL-DP-8538 aacaauagagagaaugguxTT 1207 xaccauucucucuauuguuTT 1208 NP
AL-DP-8539 ugaugauxuggcauuccaaTT 1209 uuggaaugccaxaucaucaTT 1210 NP
AL-DP-8540 augugxucucugaugcaagTT 1211 cuugcaucagagaxcacauTT 1212 NP
AL-DP-8541 ugugxucucugaugcaaggTT 1213 ccuugcaucagagaxcacaTT 1214 NP
AL-DP-8542 agxauugcauaugagagaaTT 1215 uucucucauaugcaauxcuTT 1216 NP
AL-DP-8543 gxauugcauaugagagaauTT 1217 auucucucauaugcaauxcTT 1218 NP
AL-DP-8544 xauugcauaugagagaaugTT 1219 cauucucucauaugcaauxTT 1220 NP
AL-DP-8545 ugcxuaugagagaaugugcTT 1221 gcacauucucucauaxgcaTT 1222
NP
AL-DP-8546 auaugagagaaugugcaaxTT 1223 xuugcacauucucucauauTT 1224 NP
AL-DP-8547 uaugaxagaaugugcaacaTT 1225 uguugcacauucuxucauaTT 1226 NP
AL-DP-8548 augaxagaaugugcaacauTT 1227 auguugcacauucuxucauTT 1228 NP
AL-DP-8549 aaugugcaaxauccucaaaTT 1229 uuugaggauxuugcacauuTT 1230 NP
AL-DP-8550 augugcaaxauccucaaagTT 1231 cuuugaggauxuugcacauTT 1232 NP
AL-DP-8551 ugugcaaxauccucaaaggTT 1233 ccuuugaggauxuugcacaTT 1234 NP
AL-DP-8552 agggaaauuxcaaacagcaTT 1235 ugcuguuugxaauuucccuTT 1236 NP
AL-DP-8553 gggaaauuxcaaacagcagTT 1237 cugcuguuugxaauuucccTT 1238 NP
AL-DP-8554 ggaaauuxcaaacagcagcTT 1239 gcugcuguuugxaauuuccTT 1240 NP
AL-DP-8555 gaaauuxcaaacagcagcaTT 1241 ugcugcuguuugxaauuucTT 1242 NP
AL-DP-8556 aaauuxcaaacagcagcacTT 1243 gugcugcuguuugxaauuuTT 1244 NP
AL-DP-8557 aauuxcaaacagcagcacaTT 1245 ugugcugcuguuugxaauuTT 1246 NP
AL-DP-8558 auuxcaaacagcagcacaaTT 1247 uugugcugcuguuugxaauTT 1248 NP
AL-DP-8559 gcacaaxgagcaaugauggTT 1249 ccaucauugcucxuugugcTT 1250 NP
AL-DP-8560 cacaaxgagcaaugauggaTT 1251 uccaucauugcucxuugugTT 1252 NP
AL-DP-8561 acauucccxuauacuggagTT 1253 cuccaguauaxgggaauguTT 1254
PB1 AL-DP-8562 cauucccxuauacuggagaTT 1255 ucuccaguauaxgggaaugTT
1256 PB1 AL-DP-8563 ggagaxccuccauacagccTT 1257
ggcuguauggaggxucuccTT 1258 PB1 AL-DP-8564 gagaxccuccauacagccaTT
1259 uggcuguauggaggxucucTT 1260 PB1 AL-DP-8565
agaxccuccauacagccauTT 1261 auggcuguauggaggxucuTT 1262 PB1
AL-DP-8566 gaxccuccauacagccaugTT 1263 cauggcuguauggaggxucTT 1264
PB1 AL-DP-8567 ccxccauacagccauggaaTT 1265 uuccauggcuguauggxggTT
1266 PB1 AL-DP-8568 cxccauacagccauggaacTT 1267
guuccauggcuguauggxgTT 1268 PB1 AL-DP-8569 xccauacagccauggaacaTT
1269 uguuccauggcuguauggxTT 1270 PB1 AL-DP-8570
auacagccauggaacaggxTT 1271 xccuguuccauggcuguauTT 1272 PB1
AL-DP-8571 uggaacaggxacaggauacTT 1273 guauccuguxccuguuccaTT 1274
PB1 AL-DP-8572 ggaacaggxacaggauacaTT 1275 uguauccuguxccuguuccTT
1276 PB1 AL-DP-8573 gaacaggxacaggauacacTT 1277
guguauccuguxccuguucTT 1278 PB1 AL-DP-8574 aacaggxacaggauacaccTT
1279 gguguauccuguxccuguuTT 1280 PB1 AL-DP-8575
acaggxacaggauacaccaTT 1281 ugguguauccuguxccuguTT 1282 PB1
AL-DP-8576 caggxacaggauacaccauTT 1283 augguguauccuguxccugTT 1284
PB1 AL-DP-8577 aggxacaggauacaccaugTT 1285 caugguguauccuguxccuTT
1286 PB1 AL-DP-8578 ggxacaggauacaccauggTT 1287
ccaugguguauccuguxccTT 1288 PB1 AL-DP-8579 gxacaggauacaccauggaTT
1289 uccaugguguauccuguxcTT 1290 PB1 AL-DP-8580
xacaggauacaccauggacTT 1291 guccaugguguauccuguxTT 1292 PB1
AL-DP-8581 aggauacaccauggacacxTT 1293 xguguccaugguguauccuTT 1294
PB1 AL-DP-8582 acacxgucaacagaacacaTT 1295 uguguucuguugacxguguTT
1296 PB1 AL-DP-8583 guxuuggaagcaauggcuuTT 1297
aagccauugcuuccaaxacTT 1298 PB1 AL-DP-8584 uxuuggaagcaauggcuuuTT
1299 aaagccauugcuuccaaxaTT 1300 PB1 AL-DP-8585
xuuggaagcaauggcuuucTT 1301 gaaagccauugcuuccaaxTT 1302 PB1
AL-DP-8586 gcaauggcuuuccuugaaxTT 1303 xuucaaggaaagccauugcTT 1304
PB1 AL-DP-8587 caauggcxuuccuugaagaTT 1305 ucuucaaggaaxgccauugTT
1306 PB1 AL-DP-8588 ugaaaacucxugucuugaaTT 1307
uucaagacaxgaguuuucaTT 1308 PB1 AL-DP-8589 gaaaacucxugucuugaaaTT
1309 uuucaagacaxgaguuuucTT 1310 PB1 AL-DP-8590
aaaacucxugucuugaaacTT 1311 guuucaagacaxgaguuuuTT 1312 PB1
AL-DP-8591 cgccagacxuaugacuggaTT 1313 uccagucauaxgucuggcgTT 1314
PB1 AL-DP-8592 gccagacxuaugacuggacTT 1315 guccagucauaxgucuggcTT
1316 PB1 AL-DP-8593 ccagacxuaugacuggacaTT 1317
uguccagucauaxgucuggTT 1318 PB1 AL-DP-8594 caugaccaaxaaaauggucTT
1319 gaccauuuuxuuggucaugTT 1320 PB1 AL-DP-8595
augaccaaxaaaauggucaTT 1321 ugaccauuuuxuuggucauTT 1322 PB1
AL-DP-8596 ugaccaaxaaaauggucacTT 1323 gugaccauuuuxuuggucaTT 1324
PB1 AL-DP-8597 gaccaaxaaaauggucacaTT 1325 ugugaccauuuuxuuggucTT
1326 PB1 AL-DP-8598 accaaxaaaauggucacacTT 1327
gugugaccauuuuxuugguTT 1328 PB1 AL-DP-8599 ccaaxaaaauggucacacaTT
1329 ugugugaccauuuuxuuggTT 1330 PB1 AL-DP-8600
caaxaaaauggucacacaaTT 1331 uugugugaccauuuuxuugTT 1332 PB1
AL-DP-8601 axaaaauggucacacaaagTT 1333 cuuugugugaccauuuuxuTT 1334
PB1 AL-DP-8602 ugacaxugaacacaaugacTT 1335 gucauuguguucaxugucaTT
1336 PB1 AL-DP-8603 aaxaugaugacuaacucacTT 1337
gugaguuagucaucauxuuTT 1338 PB1 AL-DP-8604 axaugaugacuaacucacaTT
1339 ugugaguuagucaucauxuTT 1340 PB1 AL-DP-8605
xaugaugacuaacucacaaTT 1341 uugugaguuagucaucauxTT 1342 PB1
AL-DP-8606 gaugacuaacucacaagaxTT 1343 xucuugugaguuagucaucTT 1344
PB1 AL-DP-8607 gacaaxaccaaauggaaugTT 1345 cauuccauuugguxuugucTT
1346 PB1 AL-DP-8608 acaaxaccaaauggaaugaTT 1347
ucauuccauuugguxuuguTT 1348 PB1 AL-DP-8609 caaxaccaaauggaaugagTT
1349 cucauuccauuugguxuugTT 1350 PB1 AL-DP-8610
aaxaccaaauggaaugagaTT 1351 ucucauuccauuugguxuuTT 1352 PB1
AL-DP-8611 axaccaaauggaaugagaaTT 1353 uucucauuccauuugguxuTT 1354
PB1 AL-DP-8612 xaccaaauggaaugagaauTT 1355 auucucauuccauuugguxTT
1356 PB1 AL-DP-8613 caaauggaaugagaaucaxTT 1357
xugauucucauuccauuugTT 1358 PB1 AL-DP-8614 auugcxccuauaauguucuTT
1359 agaacauuauaggxgcaauTT 1360 PB1 AL-DP-8615
uugcxccuauaauguucucTT 1361 gagaacauuauaggxgcaaTT 1362 PB1
AL-DP-8616 gcxccuauaauguucucaaTT 1363 uugagaacauuauaggxgcTT 1364
PB1 AL-DP-8617 cxccuauaauguucucaaaTT 1365 uuugagaacauuauaggxgTT
1366 PB1 AL-DP-8618 uacgxacacaaauaccagcTT 1367
gcugguauuuguguxcguaTT 1368 PB1 AL-DP-8619 cgxacacaaauaccagcagTT
1369 cugcugguauuuguguxcgTT 1370 PB1 AL-DP-8620
gxacacaaauaccagcagaTT 1371 ucugcugguauuuguguxcTT 1372 PB1
AL-DP-8621 acacaxauaccagcagaaaTT 1373 uuucugcugguauxuguguTT 1374
PB1 AL-DP-8622 cacaxauaccagcagaaauTT 1375 auuucugcugguauxugugTT
1376 PB1 AL-DP-8623 acaaauaccxgcagaaaugTT 1377
cauuucugcxgguauuuguTT 1378 PB1 AL-DP-8624 caaauaccxgcagaaaugcTT
1379 gcauuucugcxgguauuugTT 1380 PB1 AL-DP-8625
aaauaccxgcagaaaugcuTT 1381 agcauuucugcxgguauuuTT 1382 PB1
AL-DP-8626 aauaccagcxgaaaugcuuTT 1383 aagcauuucxgcugguauuTT 1384
PB1 AL-DP-8627 auaccagcxgaaaugcuugTT 1385 caagcauuucxgcugguauTT
1386 PB1 AL-DP-8628 uaccagcxgaaaugcuugcTT 1387
gcaagcauuucxgcugguaTT 1388 PB1 AL-DP-8629 accagcxgaaaugcuugcaTT
1389 ugcaagcauuucxgcugguTT 1390 PB1 AL-DP-8630
ccagcxgaaaugcuugcaaTT 1391 uugcaagcauuucxgcuggTT 1392 PB1
AL-DP-8631 agaaaauxgagaaaauaagTT 1393 cuuauuuucucxauuuucuTT 1394
PB1
[0055] TABLE-US-00008 TABLE 1H Exemplary iRNA agents for targeting
influenza virus derived from agents listed in Table 1C by
stabilization towards nucleolytic degradation by nucleotide
modifications Corre- Target sponding SEQ SEQ influ- Duplex
unmodified ID Antisense strand ID enza identifier duplex.sup.1
Sense strand sequence (5'-3') NO: sequence (5'-3') NO: gene
AL-DP-2336 AL-DP-2316 cmumumcmagaumcmgaacmggumcmumaTT 1395
umagaccguucgaucugaagTT 1396 PB2 AL-DP-2337 AL-DP-2317
cmagaumcmgaacmggumcmumaacmaTT 1397 uguumagaccguucgaucugTT 1398 PB2
AL-DP-2338 AL-DP-2318 agaumcmgaacmggumcmumaacmagTT 1399
cuguumagaccguucgaucuTT 1400 PB2 AL-DP-2339 AL-DP-2319
cmaaaaumgcmumaumaagumacmcmaTT 1401 uggumacuumaumagcmauuuugTT 1402
PB2 AL-DP-2340 AL-DP-2320 umumcmagaumcmgaacmggumcmumaaTT 1403
uumagaccguucgaucugaaTT 1404 PB2 AL-DP-2341 AL-DP-2321
umacmcmacmaumumcmcmcmumumaumacmumTT 1405 agumaumaagggaauguggumaTT
1406 PB2 AL-DP-2342 AL-DP-2327 umumumgagagagaagggumacmumTT 1407
agumacccuucucucucmaaaTT 1408 NP AL-DP-2343 AL-DP-2328
aggcmaacmgaacmcmcmgaumcmgumTT 1409 acgaucggguucguumgccuTT 1410 NP
AL-DP-2344 AL-DP-2329 ggcmaacmgaacmcmcmgaumcmgumgTT 1411
cmacgaucggguucguugccTT 1412 NP AL-DP-2345 AL-DP-2330
cmaacmgaacmcmcmgaumcmgumgcmcmTT 1413 ggcmacgaucggguucguugTT 1414 NP
AL-DP-2346 AL-DP-2331 gcmaacmgaacmcmcmgaumcmgumgcmTT 1415
gcmacgaucggguucguugcTT 1416 NP AL-DP-2347 AL-DP-2332
gaaaaggcmaacmgaacmcmcmgaTT 1417 ucggguucguumgccuuuucTT 1418 NP
AL-DP-2352 AL-DP-2348 umgcmaumgaumaaaggcmagumcmcmTT 1419
ggacugccuuumaucmaugcmaTT 1420 PB2 AL-DP-2353 AL-DP-2349
aumggggaumgaumcmggaaumaumTT 1421 aumauuccgaucmauccccmauTT 1422 PB2
AL-DP-2354 AL-DP-2350 umggggaumgaumcmggaaumaumumTT 1423
aaumauuccgaucmauccccmaTT 1424 PB2 AL-DP-2355 AL-DP-2351
gaaacmgggacmumcmumagcmaumaTT 1425 umaugcumagagucccguuucTT 1426 PB2
AL-DP-2369 AL-DP-2356 agacmumumumgumgcmgacmaaumgcmTT 1427
gcmauugucgcmacmaaagucuTT 1428 PA AL-DP-2370 AL-DP-2357
gacmumumumgumgcmgacmaaumgcmumTT 1429 agcmauugucgcmacmaaagucTT 1430
PA AL-DP-2371 AL-DP-2358 umcmumaumgggaumumcmcmumumumcmgumTT 1431
acgaaaggaaucccmaumagaTT 1432 PA AL-DP-2372 AL-DP-2359
cmumaumgggaumumcmcmumumumcmgumcmTT 1433 gacgaaaggaaucccmaumagTT
1434 PA AL-DP-2373 AL-DP-2360 aumgumggaumggaumumcmgaacmcmTT 1435
gguucgaauccmauccmacmauTT 1436 PA AL-DP-2374 AL-DP-2361
umgumggaumggaumumcmgaacmcmgTT 1437 cgguucgaauccmauccmacmaTT 1438 PA
AL-DP-2375 AL-DP-2362 gumggaumggaumumcmgaacmcmgaTT 1439
ucgguucgaauccmauccmacTT 1440 PA AL-DP-2376 AL-DP-2363
umggaumggaumumcmgaacmcmgaaTT 1441 uucgguucgaauccmauccmaTT 1442 PA
AL-DP-2377 AL-DP-2364 ggaumggaumumcmgaacmcmgaacmTT 1443
guucgguucgaauccmauccTT 1444 PA AL-DP-2378 AL-DP-2365
gaumggaumumcmgaacmcmgaacmgTT 1445 cguucgguucgaauccmaucTT 1446 PA
AL-DP-2379 AL-DP-2366 aumggaumumcmgaacmcmgaacmggTT 1447
ccguucgguucgaauccmauTT 1448 PA AL-DP-2380 AL-DP-2367
umggaumumcmgaacmcmgaacmggcmTT 1449 gccguucgguucgaauccmaTT 1450 PA
AL-DP-2381 AL-DP-2368 aumcmumcmcmacmaacmumcmgaggggTT 1451
ccccucgaguumgumggagauTT 1452 PA .sup.1duplex identifier of siRNA
agent of Table 1C having an identical nucleotide sequence when
nucleotide modifications are disregarded
[0056] TABLE-US-00009 TABLE 1I Activity of the modified RNAi agents
listed in Table 1B and H towards inhibition of influenza gene
expression in the assays described in Example 3 Target ELISA (MDCK
ELISA (Vero Plasmid Duplex influenza % remaining cells), cells),
expression, identifier gene infectivity.sup.1 % inhibition.sup.2 %
inhibition.sup.3 % inhibition.sup.4 AL-DP-2289 PB1 104% 17 -31
AL-DP-2290 NP 29% -8 -36 61 AL-DP-2291 NP 34% -28 -30 5 AL-DP-2292
NP 34% -25 -14 36 AL-DP-2293 MP 40% -7 -74 AL-DP-2294 MP 78% -19
-53 AL-DP-2295 MP 67% -39 -85 AL-DP-2296 MP 61% -21 AL-DP-2297 MP
-15 -27 AL-DP-2298 MP -21 11 AL-DP-2299 MP -23 12 AL-DP-2300 MP -37
-62 AL-DP-2301 MP -13 -62 AL-DP-2302 MP -30 -51 AL-DP-2303 MP 1 -44
AL-DP-2304 MP -16 -38 AL-DP-2305 MP 45% 28 -42 AL-DP-2306 MP 46% -1
-46 AL-DP-2307 MP 39% 11 -18 AL-DP-2308 MP -5 15 AL-DP-2309 MP 19
-42 AL-DP-2310 MP -1 -29 AL-DP-2311 MP -46 -45 AL-DP-2312 MP -66
-31 AL-DP-2336 PB2 11 -15 AL-DP-2337 PB2 6 -23 AL-DP-2338 PB2 5 5
AL-DP-2339 PB2 33 -38 AL-DP-2340 PB2 19 -46 AL-DP-2341 PB2 14 -5
AL-DP-2342 NP 9 3 42 AL-DP-2343 NP -32 -20 29 AL-DP-2344 NP -27 -10
22 AL-DP-2345 NP 15 -29 39 AL-DP-2346 NP -22 -32 29 AL-DP-2347 NP
-9 -24 65 AL-DP-2352 PB2 3 17 5 AL-DP-2353 PB2 -44 9 28 AL-DP-2354
PB2 -54 -9 27 AL-DP-2355 PB2 <25 13 45 59 AL-DP-2369 PA <75
40 3 2 AL-DP-2370 PA 28 27 67 AL-DP-2371 PA 15 -24 12 AL-DP-2372 PA
18 -29 73 AL-DP-2373 PA <25 37 27 71 AL-DP-2374 PA <75 27 -48
9 AL-DP-2375 PA <25 44 53 87 AL-DP-2376 PA 4 -40 38 AL-DP-2377
PA <25 21 39 65 AL-DP-2378 PA -50 -75 11 AL-DP-2379 PA -39 -20
19 AL-DP-2380 PA 0 -27 31 AL-DP-2381 PA -52 -54 43 .sup.1in in
vitro plaque forming assay in MCDK cells as described in Example
3.1; .sup.2in in vitro ELISA assay in MCDK cells as described in
Example 3.2: .sup.3in in vitro ELISA assay in MCDK cells as
described in Example 3.2; .sup.4in in vitro ELISA assay in MCDK
cells as described in Example 3.2; negative values indicate that
target gene expression was enhanced in treated cells compared to
controls
[0057] TABLE-US-00010 TABLE 2 Concentration at 50% inhibition
(IC.sub.50) for selected RNAi agents of Table 1 Target Duplex
influenza identifier gene IC.sub.50 (nM) AL-DP-2364 PA 0.22
AL-DP-2377 PA 2.15 AL-DP-7595 PB2 0.54 AL-DP-7611 PB2 0.075
AL-DP-7617 NP 0.57 AL-DP-7622 NP 0.74 AL-DP-7633 NP .about.90
AL-DP-7660 M 261 AL-DP-7669 M 0.79
[0058] The antisense strand of an iRNA agent should be equal to or
at least, 14, 15, 16 17, 18, 19, 25, 29, 40, or 50 nucleotides in
length. It should be equal to or less than 60, 50, 40, or 30,
nucleotides in length. Preferred ranges are 15-30, 17 to 25, 19 to
23, and 19 to 21 nucleotides in length.
[0059] The sense strand of an iRNA agent should be equal to or at
least 14, 15, 16 17, 18, 19, 25, 29, 40, or 50 nucleotides in
length. It should be equal to or less than 60, 50, 40, or 30
nucleotides in length. Preferred ranges are 15-30, 17 to 25, 19 to
23, and 19 to 21 nucleotides in length.
[0060] The double stranded portion of an iRNA agent should be equal
to or at least, 15, 16 17, 18, 19, 20, 21, 22, 23, 24, 25, 29, 40,
or 50 nucleotide pairs in length. It should be equal to or less
than 60, 50, 40, or 30 nucleotides pairs in length. Preferred
ranges are 15-30, 17 to 25, 19 to 23, and 19 to 21 nucleotides
pairs in length.
[0061] Generally, the iRNA agents of the instant invention include
a region of sufficient complementarity to the respective influenza
virus gene, and are of sufficient length in terms of nucleotides,
that the iRNA agent, or a fragment thereof, can mediate down
regulation of the influenza virus gene. It is not necessary that
there be perfect complementarity between the iRNA agent and the
target gene, but the correspondence must be sufficient to enable
the iRNA agent, or a cleavage product thereof, to direct sequence
specific silencing, e.g., by RNAi cleavage of an influenza virus
RNA.
[0062] Therefore, the iRNA agents of the instant invention include
agents comprising a sense strand and antisense strand each
comprising a sequence of at least 16, 17 or 18 nucleotides which is
essentially identical, as defined below, to one of the sequences of
Tables 1A-1H, except that not more than 1, 2 or 3 nucleotides per
strand, respectively, have been substituted by other nucleotides
(e.g. adenosine replaced by uracil), while essentially retaining
the ability to inhibit influenza virus replication in cultured
human cells infected with influenza virus, respectively. These
agents will therefore possess at least 15 nucleotides identical to
one of the sequences of Tables 1A-1H, but 1, 2 or 3 base mismatches
with respect to either the target influenza virus RNA sequence or
between the sense and antisense strand are introduced. Mismatches
to the target influenza virus RNA sequence, particularly in the
antisense strand, are most tolerated in the terminal regions and if
present are preferably in a terminal region or regions, e.g.,
within 6, 5, 4, or 3 nucleotides of a 5' and/or 3' terminus, most
preferably within 6, 5, 4, or 3 nucleotides of the 5'-terminus of
the sense strand or the 3'-terminus of the antisense strand. The
sense strand need only be sufficiently complementary with the
antisense strand to maintain the overall double stranded character
of the molecule.
[0063] It is preferred that the sense and antisense strands be
chosen such that the iRNA agent includes a single strand or
unpaired region at one or both ends of the molecule. Thus, an iRNA
agent contains sense and antisense strands, preferably paired to
contain an overhang, e.g., one or two 5' or 3' overhangs but
preferably a 3' overhang of 2-3 nucleotides. Most embodiments will
have a 3' overhang. Preferred siRNA agents will have
single-stranded overhangs, preferably 3' overhangs, of 1 to 4, or
preferably 2 or 3 nucleotides, in length, at one or both ends of
the iRNA agent. The overhangs can be the result of one strand being
longer than the other, or the result of two strands of the same
length being staggered. The unpaired nucleotides forming the
overhang can be ribonucleotides, or they can be
deoxyribonucleotides, preferably thymidine. 5'-ends are preferably
phosphorylated, or they may be unphosphorylated.
[0064] Preferred lengths for the duplexed region are between 15 and
30, most preferably 18, 19, 20, 21, 22, and 23 nucleotides in
length, e.g., in the siRNA agent range discussed above. siRNA
agents can resemble in length and structure the natural Dicer
processed products from long dsRNAs. Embodiments in which the two
strands of the siRNA agent are linked, e.g., covalently linked, are
also included. Hairpin, or other single strand structures which
provide the required double stranded region, and preferably a 3'
overhang are also within the invention.
[0065] Evaluation of Candidate iRNA Agents
[0066] As noted above, the present invention provides a system for
identifying siRNAs that are useful as inhibitors of influenza virus
infection and/or replication. Since, as noted above, shRNAs are
processed intracellularly to produce siRNAs having duplex portions
with the same sequence as the stem structure of the shRNA, the
system is equally useful for identifying shRNAs that are useful as
inhibitors of influenza virus infection. For purposes of
description this section will refer to siRNAs, but the system also
encompasses corresponding shRNAs. Specifically, the present
invention demonstrates the successful preparation of siRNAs
targeted to viral genes to block or inhibit viral infection and/or
replication. The techniques and reagents described herein can
readily be applied to design potential new siRNAs, targeted to
other genes or gene regions, and tested for their activity in
inhibiting influenza virus infection and/or replication as
discussed herein. It is expected that influenza viruses will
continue to mutate and undergo reassortment and that it may be
desirable to continue to develop and test new, differently targeted
siRNAs.
[0067] In various embodiments of the invention potential influenza
virus inhibitors can be tested by introducing candidate siRNA(s)
into cells (e.g., by exogenous administration or by introducing a
vector or construct that directs endogenous synthesis of siRNA into
the cell), or laboratory animals, prior to, simultaneously with, or
after transfection with an influenza genome or portion thereof
(e.g., within minutes, hours, or at most a few days) or prior to,
simultaneously with, or after infection with influenza virus.
Alternately, potential influenza virus inhibitors can be tested by
introducing candidate siRNA(s) into cells or laboratory animals
that are productively infected with influenza virus (i.e., cells
that are producing progeny virus). The ability of the candidate
siRNA(s) to reduce target transcript levels and/or to inhibit or
suppress one or more aspects or features of the viral life cycle
such as viral replication, pathogenicity, and/or infectivity is
then assessed. For example, production of viral particles and/or
production of viral proteins, etc., can be assessed either directly
or indirectly using methods well known in the art.
[0068] Cells or laboratory animals to which inventive siRNA
compositions have been delivered (test cells/animals) may be
compared with similar or comparable cells or laboratory animals
that have not received the inventive composition (control
cells/animals, e.g., cells/animals that have received either no
siRNA or a control siRNA such as an siRNA targeted to a non-viral
transcript such as GFP). The susceptibility of the test
cells/animals to influenza virus infection can be compared with the
susceptibility of control cells/animals to infection. Production of
viral protein(s) and/or progeny virus may be compared in the test
cells/animals relative to the control cells/animals. Other indicia
of viral infectivity, replication, pathogenicity, etc., can be
similarly compared. Standard in vitro antiviral assays may utilize
inhibition of viral plaques, viral cytopathic effect (CPE), and
viral hemagglutinin or other protein, inhibition of viral yield,
etc. The CPE can be determined visually and by dye uptake. See,
e.g., Sidwell, R. W. and Smee, D. F, "In vitro and in vivo assay
systems for study of influenza virus inhibitors" Antiviral Res
2000, 48:1. Generally, test cells/animals and control cells/animals
would be from the same species and, for cells, of similar or
identical cell type. For example, cells from the same cell line
could be compared. When the test cell is a primary cell, typically
the control cell would also be a primary cell. Typically the same
influenza virus strain would be used to compare test cells/animals
and control cells/animals.
[0069] For example, the ability of a candidate siRNA to inhibit
influenza virus production may conveniently be determined by (i)
delivering the candidate siRNA to cells (either prior to, at the
same time as, or after exposure to influenza virus); (ii) assessing
the production of viral hemagglutinin using a hemagglutinin assay,
and (iii) comparing the amount of hemagglutinin produced in the
presence of the siRNA with the amount produced in the absence of
the siRNA. (The test need not include a control in which the siRNA
is absent but may make use of previous information regarding the
amount of hemagglutinin produced in the absence of inhibition.) A
reduction in the amount of hemagglutinin strongly suggests a
reduction in virus production. This assay may be used to test
siRNAs that target any viral transcript and is not limited to
siRNAs that target the transcript that encodes the viral
hemagglutinin.
[0070] The ability of a candidate siRNA to reduce the level of the
target transcript may also be assessed by measuring the amount of
the target transcript using, for example, Northern blots, nuclease
protection assays, reverse transcription (RT)-PCR, real-time
RT-PCR, microarray analysis, etc. The ability of a candidate siRNA
to inhibit production of a polypeptide encoded by the target
transcript (either at the transcriptional or post-transcriptional
level) may be measured using a variety of antibody-based approaches
including, but not limited to, Western blots, immunoassays, ELISA,
flow cytometry, protein microarrays, etc. In general, any method of
measuring the amount of either the target transcript or a
polypeptide encoded by the target transcript may be used.
[0071] In general, certain preferred influenza virus inhibitors
reduce the target transcript level at least about 2 fold,
preferably at least about 4 fold, more preferably at least about 8
fold, at least about 16 fold, at least about 64 fold or to an even
greater degree relative to the level that would be present in the
absence of the inhibitor (e.g., in a comparable control cell
lacking the inhibitor). In general, certain preferred influenza
virus inhibitors inhibit viral replication, so that the level of
replication is lower in a cell containing the inhibitor than in a
control cell not containing the inhibitor by at least about 2 fold,
preferably at least about 4 fold, more preferably at least about 8
fold, at least about 16 fold, at least about 64 fold, at least
about 100 fold, at least about 200 fold, or to an even greater
degree.
[0072] Certain preferred influenza virus inhibitors inhibit viral
replication so that development of detectable viral titer is
prevented for at least 24 hours, at least 36 hours, at least 48
hours, or at least 60 hours following administration of the siRNA
and infection of the cells. Certain preferred influenza virus
inhibitors prevent (i.e., reduce to undetectable levels) or
significantly reduce viral replication for at least 24 hours, at
least 36 hours, at least 48 hours, or at least 60 hours following
administration of the siRNA. According to various embodiments of
the invention a significant reduction in viral replication is a
reduction to less than approximately 90% of the level that would
occur in the absence of the siRNA, a reduction to less than
approximately 75% of the level that would occur in the absence of
the siRNA, a reduction to less than approximately 50% of the level
that would occur in the absence of the siRNA, a reduction to less
than approximately 25% of the level that would occur in the absence
of the siRNA, or a reduction to less than approximately 10% of the
level that would occur in the absence of the siRNA. Reduction in
viral replication may be measured using any suitable method
including, but not limited to, measurement of HA titer.
[0073] Stability Testing, Modification, and Retesting of iRNA
Agents
[0074] A candidate iRNA agent can be evaluated with respect to
stability, e.g., its susceptibility to cleavage by an endonuclease
or exonuclease, such as when the iRNA agent is introduced into the
body of a subject. Methods can be employed to identify sites that
are susceptible to modification, particularly cleavage, e.g.,
cleavage by a component found in the body of a subject. Such
methods may include the isolation and identification of most
abundant fragments formed by degradation of the candidate iRNA
agent after its incubation with isolated biological media in vitro,
e.g. serum, plasma, sputum, cerebrospinal fluid, or cell or tissue
homogenates, or after contacting a subject with the candidate iRNA
agent in vivo, thereby identifying sites prone to cleavage. Such
methods are, for example, without limitation, in co-owned
International Application No. PCT/US2005/018931, filed on May 27,
2005.
[0075] When sites susceptible to cleavage are identified, a further
iRNA agent can be designed and/or synthesized wherein the potential
cleavage site is made resistant to cleavage, e.g. by introduction
of a 2'-modification on the site of cleavage, e.g. a 2'-O-methyl
group. This further iRNA agent can be retested for stability, and
this process may be iterated until an iRNA agent is found
exhibiting the desired stability.
[0076] In Vivo Testing
[0077] An iRNA agent identified as being capable of inhibiting
influenza virus gene expression can be tested for functionality in
vivo in an animal model (e.g., in a mammal, such as in mouse or
rat). For example, the iRNA agent can be administered to an animal,
and the iRNA agent evaluated with respect to its biodistribution,
stability, and its ability to inhibit influenza virus replication
or reduce a biological or pathological process mediated at least in
part by influenza virus.
[0078] The iRNA agent can be administered directly to the target
tissue, such as by injection, or the iRNA agent can be administered
to the animal model in the same manner that it would be
administered to a human. Preferably, the iRNA agent is delivered to
the subject's airways, such as intranasally.
[0079] The iRNA agent can also be evaluated for its intracellular
distribution. The evaluation can include determining whether the
iRNA agent was taken up into the cell. The evaluation can also
include determining the stability (e.g., the half-life) of the iRNA
agent. Evaluation of an iRNA agent in vivo can be facilitated by
use of an iRNA agent conjugated to a traceable marker (e.g., a
fluorescent marker such as fluorescein; a radioactive label, such
as .sup.35S, .sup.32P, .sup.33P, or .sup.3H; gold particles; or
antigen particles for immunohistochemistry).
[0080] The iRNA agent can be evaluated with respect to its ability
to down regulate influenza virus replication. Levels of influenza
virus gene expression in vivo can be measured, for example, by in
situ hybridization, or by the isolation of RNA from tissue prior to
and following exposure to the iRNA agent. Where the animal needs to
be sacrificed in order to harvest the tissue, an untreated control
animal will serve for comparison. Influenza virus RNA can be
detected by any desired method, including but not limited to
RT-PCR, Northern blot, branched-DNA assay, or RNAase protection
assay. Alternatively, or additionally, influenza virus gene
expression can be monitored by performing Western blot analysis or
plaque forming assays on tissue extracts treated with the iRNA
agent.
[0081] Potential influenza virus inhibitors can be tested using any
of variety of animal models that have been developed. Compositions
comprising candidate siRNA(s), constructs or vectors capable of
directing synthesis of such siRNAs within a host cell, or cells
engineered or manipulated to contain candidate siRNAs may be
administered to an animal prior to, simultaneously with, or
following infection with an influenza virus. The ability of the
composition to prevent viral infection and/or to delay or prevent
appearance of influenza-related symptoms and/or lessen their
severity relative to influenza-infected animals that have not
received the potential influenza inhibitor is assessed. Such models
include, but are not limited to, murine, chicken, ferret, and
non-human primate models for influenza infection, all of which are
known in the art and are used for testing the efficacy of potential
influenza therapeutics and vaccines. See, e.g, Sidwell, R. W. and
Smee, D. F, referenced above. Such models may involve use of
naturally occurring influenza virus strains and/or strains that
have been modified or adapted to existence in a particular host
(e.g., the WSN or PR8 strains, which are adapted for replication in
mice). The above animal models may also be used to establish the
concentration necessary to achieve a certain desired effect (e.g.,
EC50).
[0082] IRNA Chemistry
[0083] Described herein are isolated iRNA agents, e.g., ds RNA
agents that mediate RNAi to inhibit expression of a influenza virus
gene.
[0084] RNA agents discussed herein include otherwise unmodified RNA
as well as RNA which has been modified, e.g., to improve efficacy,
and polymers of nucleoside surrogates. Unmodified RNA refers to a
molecule in which the components of the nucleic acid, namely
sugars, bases, and phosphate moieties, are the same or essentially
the same as that which occur in nature, preferably as occur
naturally in the human body. The art has referred to rare or
unusual, but naturally occurring, RNAs as modified RNAs, see, e.g.,
Limbach et al. Nucleic Acids Res. 22: 2183-2196, 1994. Such rare or
unusual RNAs, often termed modified RNAs (apparently because they
are typically the result of a post-transcriptional modification)
are within the term unmodified RNA, as used herein. Modified RNA as
used herein refers to a molecule in which one or more of the
components of the nucleic acid, namely sugars, bases, and phosphate
moieties, are different from that which occurs in nature,
preferably different from that which occurs in the human body.
While they are referred to as modified "RNAs," they will of course,
because of the modification, include molecules which are not RNAs.
Nucleoside surrogates are molecules in which the ribophosphate
backbone is replaced with a non-ribophosphate construct that allows
the bases to the presented in the correct spatial relationship such
that hybridization is substantially similar to what is seen with a
ribophosphate backbone, e.g., non-charged mimics of the
ribophosphate backbone. Examples of the above are discussed
herein.
[0085] Modifications described herein can be incorporated into any
double-stranded RNA and RNA-like molecule described herein, e.g.,
an iRNA agent. It may be desirable to modify one or both of the
antisense and sense strands of an iRNA agent. As nucleic acids are
polymers of subunits or monomers, many of the modifications
described below occur at a position which is repeated within a
nucleic acid, e.g., a modification of a base, or a phosphate
moiety, or the non-linking 0 of a phosphate moiety. In some cases
the modification will occur at all of the subject positions in the
nucleic acid but in many, and in fact in most, cases it will not.
By way of example, a modification may only occur at a 3' or 5'
terminal position, may only occur in a terminal region, e.g. at a
position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10
nucleotides of a strand. A modification may occur in a double
strand region, a single strand region, or in both. E.g., a
phosphorothioate modification at a non-linking O position may only
occur at one or both termini, may only occur in a terminal regions,
e.g., at a position on a terminal nucleotide or in the last 2, 3,
4, 5, or 10 nucleotides of a strand, or may occur in double strand
and single strand regions, particularly at termini. Similarly, a
modification may occur on the sense strand, antisense strand, or
both. In some cases, the sense and antisense strand will have the
same modifications or the same class of modifications, but in other
cases the sense and antisense strand will have different
modifications, e.g., in some cases it may be desirable to modify
only one strand, e.g. the sense strand.
[0086] Two prime objectives for the introduction of modifications
into iRNA agents is their stabilization towards degradation in
biological environments and the improvement of pharmacological
properties, e.g. pharmacodynamic properties, which are further
discussed below. Other suitable modifications to a sugar, base, or
backbone of an iRNA agent are described in co-owned PCT Application
No. PCT/US2004/01193, filed Jan. 16, 2004. An iRNA agent can
include a non-naturally occurring base, such as the bases described
in co-owned PCT Application No. PCT/US2004/011822, filed Apr. 16,
2004. An iRNA agent can include a non-naturally occurring sugar,
such as a non-carbohydrate cyclic carrier molecule. Exemplary
features of non-naturally occurring sugars for use in iRNA agents
are described in co-owned PCT Application No. PCT/US2004/11829,
filed Apr. 16, 2003.
[0087] An iRNA agent can include an internucleotide linkage (e.g.,
the chiral phosphorothioate linkage) useful for increasing nuclease
resistance. In addition, or in the alternative, an iRNA agent can
include a ribose mimic for increased nuclease resistance. Exemplary
internucleotide linkages and ribose mimics for increased nuclease
resistance are described in co-owned PCT Application No.
PCT/US2004/07070, filed on Mar. 8, 2004.
[0088] An iRNA agent can include ligand-conjugated monomer subunits
and monomers for oligonucleotide synthesis. Exemplary monomers are
described in co-owned U.S. application Ser. No. 10/916,185, filed
on Aug. 10, 2004.
[0089] An iRNA agent can have a ZXY structure, such as is described
in co-owned PCT Application No. PCT/US2004/07070, filed on Mar. 8,
2004.
[0090] An iRNA agent can be complexed with an amphipathic moiety.
Exemplary amphipathic moieties for use with iRNA agents are
described in co-owned PCT Application No. PCT/US2004/07070, filed
on Mar. 8, 2004.
[0091] In another embodiment, the iRNA agent can be complexed to a
delivery agent that features a modular complex. The complex can
include a carrier agent linked to one or more of (preferably two or
more, more preferably all three of): (a) a condensing agent (e.g.,
an agent capable of attracting, e.g., binding, a nucleic acid,
e.g., through ionic or electrostatic interactions); (b) a fusogenic
agent (e.g., an agent capable of fusing and/or being transported
through a cell membrane); and (c) a targeting group, e.g., a cell
or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or
protein, e.g., an antibody, that binds to a specified cell type.
iRNA agents complexed to a delivery agent are described in co-owned
PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004.
[0092] An iRNA agent can have non-canonical pairings, such as
between the sense and antisense sequences of the iRNA duplex.
Exemplary features of non-canonical iRNA agents are described in
co-owned PCT Application No. PCT/US2004/07070, filed on Mar. 8,
2004.
[0093] Enhanced Nuclease Resistance
[0094] An iRNA agent, e.g., an iRNA agent that targets influenza
virus, can have enhanced resistance to nucleases.
[0095] One way to increase resistance is to identify cleavage sites
and modify such sites to inhibit cleavage, as described in co-owned
U.S. Application No. 60/559,917, filed on May 4, 2004. For example,
the dinucleotides 5'-ua-3',5'-ca-3',5'-ug-3',5'-uu-3', or 5'-ca-3'
can serve as cleavage sites. In certain embodiments, all the
pyrimidines of an iRNA agent carry a 2'-modification in either the
sense strand, the antisense strand, or both strands, and the iRNA
agent therefore has enhanced resistance to endonucleases. Enhanced
nuclease resistance can also be achieved by modifying the 5'
nucleotide, resulting, for example, in at least one
5'-uridine-adenine-3' (5'-ua-3') dinucleotide wherein the uridine
is a 2'-modified nucleotide; at least one 5'-cytidine-adenine-3'
(5'-ca-3') dinucleotide, wherein the 5'-cytidine is a 2'-modified
nucleotide; at least one 5'-uridine-guanine-3' (5'-ug-3')
dinucleotide, wherein the 5'-uridine is a 2'-modified nucleotide;
at least one 5'-uridine-uridine-3' (5'-uu-3') dinucleotide, wherein
the 5'-uridine is a 2'-modified nucleotide; or at least one
5'-cytidine-cytidine-3' (5'-ca-3') dinucleotide, wherein the
5'-cytidine is a 2'-modified nucleotide, as described in co-owned
International Application No. PCT/US2005/018931, filed on May 27,
2005. The iRNA agent can include at least 2, at least 3, at least 4
or at least 5 of such dinucleotides. In a particularly preferred
embodiment, the 5' nucleotide in all occurrences of the sequence
motifs 5'-ua-3' and 5'-ca-3' in either the sense strand, the
antisense strand, or both strands is a modified nucleotide.
Preferably, the 5' nucleotide in all occurrences of the sequence
motifs 5'-ua-3',5'-ca-3' and 5'-ug-3' in either the sense strand,
the antisense strand, or both strands is a modified nucleotide.
More preferably, all pyrimidine nucleotides in the sense strand are
modified nucleotides, and the 5' nucleotide in all occurrences of
the sequence motifs 5'-ua-3' and 5'-ca-3' in the antisense strand
are modified nucleotides, or where the antisense strand does
comprise neither of a 5'-ua-3' and a 5'-ca-3' motif, in all
occurrences of the sequence motif 5'-ug-3'.
[0096] Preferably, the 2'-modified nucleotides include, for
example, a 2'-modified ribose unit, e.g., the 2'-hydroxyl group
(OH) can be modified or replaced with a number of different "oxy"
or "deoxy" substituents.
[0097] Examples of "oxy"-2' hydroxyl group modifications include
alkoxy or aryloxy (OR, e.g., R.dbd.H, alkyl, cycloalkyl, aryl,
aralkyl, heteroaryl or sugar); polyethyleneglycols (PEG),
O(CH.sub.2CH.sub.2O).sub.nCH.sub.2CH.sub.2OR; "locked" nucleic
acids (LNA) in which the 2' hydroxyl is connected, e.g., by a
methylene bridge, to the 4' carbon of the same ribose sugar;
O-AMINE and aminoalkoxy, O(CH.sub.2).sub.nAMINE, (e.g.,
AMINE=NH.sub.2; alkylamino, dialkylamino, heterocyclyl amino,
arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino,
ethylene diamine, polyamino). It is noteworthy that
oligonucleotides containing only the methoxyethyl group (MOE),
(OCH.sub.2CH.sub.2OCH.sub.3, a PEG derivative), exhibit nuclease
stabilities comparable to those modified with the robust
phosphorothioate modification.
[0098] "Deoxy" modifications include hydrogen (i.e. deoxyribose
sugars, which are of particular relevance to the overhang portions
of partially ds RNA); halo (e.g., fluoro); amino (e.g. NH.sub.2;
alkylamino, dialkylamino, heterocyclyl, arylamino, diaryl amino,
heteroaryl amino, diheteroaryl amino, or amino acid);
NH(CH.sub.2CH.sub.2NH).sub.nCH.sub.2CH.sub.2-AMINE (AMINE=NH.sub.2;
alkylamino, dialkylamino, heterocyclyl amino, arylamino, diaryl
amino, heteroaryl amino, or diheteroaryl amino), --NHC(O)R
(R=alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar), cyano;
mercapto; alkyl-thio-alkyl; thioalkoxy; and alkyl, cycloalkyl,
aryl, alkenyl and alkynyl, which may be optionally substituted with
e.g., an amino functionality.
[0099] Preferred substitutents are 2'-methoxyethyl, 2'-OCH.sub.3,
2'-O-allyl, 2'-C-allyl, and 2'-fluoro.
[0100] The inclusion of furanose sugars in the oligonucleotide
backbone can also decrease endonucleolytic cleavage. An iRNA agent
can be further modified by including a 3' cationic group, or by
inverting the nucleoside at the 3'-terminus with a 3'-3' linkage.
In another alternative, the 3'-terminus can be blocked with an
aminoalkyl group, e.g., a 3' C5-aminoalkyl dT. Other 3' conjugates
can inhibit 3'-5' exonucleolytic cleavage. While not being bound by
theory, a 3' conjugate, such as naproxen or ibuprofen, may inhibit
exonucleolytic cleavage by sterically blocking the exonuclease from
binding to the 3'-end of oligonucleotide. Even small alkyl chains,
aryl groups, or heterocyclic conjugates or modified sugars
(D-ribose, deoxyribose, glucose etc.) can block
3'-5'-exonucleases.
[0101] Nucleolytic cleavage can also be inhibited by the
introduction of phosphate linker modifications, e.g.,
phosphorothioate linkages. Thus, preferred iRNA agents include
nucleotide dimers enriched or pure for a particular chiral form of
a modified phosphate group containing a heteroatom at a nonbridging
position normally occupied by oxygen. The heteroatom can be S, Se,
Nr.sub.2, or Br.sub.3. When the heteroatom is S, enriched or
chirally pure Sp linkage is preferred. Enriched means at least 70,
80, 90, 95, or 99% of the preferred form. Modified phosphate
linkages are particularly efficient in inhibiting exonucleolytic
cleavage when introduced near the 5'- or 3'-terminal positions, and
preferably the 5'-terminal positions, of an iRNA agent.
[0102] 5' conjugates can also inhibit 5'-3' exonucleolytic
cleavage. While not being bound by theory, a 5' conjugate, such as
naproxen or ibuprofen, may inhibit exonucleolytic cleavage by
sterically blocking the exonuclease from binding to the 5'-end of
oligonucleotide. Even small alkyl chains, aryl groups, or
heterocyclic conjugates or modified sugars (D-ribose, deoxyribose,
glucose etc.) can block 3'-5'-exonucleases.
[0103] An iRNA agent can have increased resistance to nucleases
when a duplexed iRNA agent includes a single-stranded nucleotide
overhang on at least one end. In preferred embodiments, the
nucleotide overhang includes 1 to 4, preferably 2 to 3, unpaired
nucleotides. In a preferred embodiment, the unpaired nucleotide of
the single-stranded overhang that is directly adjacent to the
terminal nucleotide pair contains a purine base, and the terminal
nucleotide pair is a G-C pair, or at least two of the last four
complementary nucleotide pairs are G-C pairs. In further
embodiments, the nucleotide overhang may have 1 or 2 unpaired
nucleotides, and in an exemplary embodiment the nucleotide overhang
is 5'-gc-3'. In preferred embodiments, the nucleotide overhang is
on the 3'-end of the antisense strand. In one embodiment, the iRNA
agent includes the motif 5'-cgc-3' on the 3'-end of the antisense
strand, such that a 2-nt overhang 5'-gc-3' is formed.
[0104] Thus, an iRNA agent can include modifications so as to
inhibit degradation, e.g., by nucleases, e.g., endonucleases or
exonucleases, found in the body of a subject. These monomers are
referred to herein as NRMs, or Nuclease Resistance promoting
Monomers, the corresponding modifications as NRM modifications. In
many cases these modifications will modulate other properties of
the iRNA agent as well, e.g., the ability to interact with a
protein, e.g., a transport protein, e.g., serum albumin, or a
member of the RISC, or the ability of the first and second
sequences to form a duplex with one another or to form a duplex
with another sequence, e.g., a target molecule.
[0105] One or more different NRM modifications can be introduced
into an iRNA agent or into a sequence of an iRNA agent. An NRM
modification can be used more than once in a sequence or in an iRNA
agent.
[0106] NRM modifications include some which can be placed only at
the terminus and others which can go at any position. Some NRM
modifications can inhibit hybridization so it is preferable to use
them only in terminal regions, and preferable to not use them at
the cleavage site or in the cleavage region of a sequence which
targets a subject sequence or gene, particularly on the antisense
strand. They can be used anywhere in a sense strand, provided that
sufficient hybridization between the two strands of the ds iRNA
agent is maintained. In some embodiments it is desirable to put the
NRM at the cleavage site or in the cleavage region of a sense
strand, as it can minimize off-target silencing.
[0107] In most cases, NRM modifications will be distributed
differently depending on whether they are comprised on a sense or
antisense strand. If on an antisense strand, modifications which
interfere with or inhibit endonuclease cleavage should not be
inserted in the region which is subject to RISC mediated cleavage,
e.g., the cleavage site or the cleavage region (As described in
Elbashir et al., 2001, Genes and Dev. 15: 188, hereby incorporated
by reference). Cleavage of the target occurs about in the middle of
a 20 or 21 nt antisense strand, or about 10 or 11 nucleotides
upstream of the first nucleotide on the target mRNA which is
complementary to the antisense strand. As used herein cleavage site
refers to the nucleotides on either side of the cleavage site, on
the target or on the iRNA agent strand which hybridizes to it.
Cleavage region means the nucleotides within 1, 2, or 3 nucleotides
of the cleavagee site, in either direction.
[0108] Such modifications can be introduced into the terminal
regions, e.g., at the terminal position or with 2, 3, 4, or 5
positions of the terminus, of a sense or antisense strand.
[0109] Tethered Ligands
[0110] The properties of an iRNA agent, including its
pharmacological properties, can be influenced and tailored, for
example, by the introduction of ligands, e.g. tethered ligands. In
addition, pharmacological properties of an iRNA agent can be
improved by incorporating a ligand in a formulation of the iRNA
agent when the iRNA agent either has or does have a tethered
ligand.
[0111] A wide variety of entities, e.g., ligands, can be tethered
to an iRNA agent or used as formulation conjugate or additive,
e.g., to the carrier of a ligand-conjugated monomer subunit.
Examples are described below in the context of a ligand-conjugated
monomer subunit but that is only preferred, entities can be coupled
at other points to an iRNA agent.
[0112] Preferred moieties are ligands, which are coupled,
preferably covalently, either directly or indirectly via an
intervening tether, to the carrier. In preferred embodiments, the
ligand is attached to the carrier via an intervening tether. The
ligand or tethered ligand may be present on the ligand-conjugated
monomer when the ligand-conjugated monomer is incorporated into the
growing strand. In some embodiments, the ligand may be incorporated
into a "precursor" ligand-conjugated monomer subunit after a
"precursor" ligand-conjugated monomer subunit has been incorporated
into the growing strand. For example, a monomer having, e.g., an
amino-terminated tether, e.g., TAP-(CH.sub.2).sub.nNH.sub.2 may be
incorporated into a growing sense or antisense strand. In a
subsequent operation, i.e., after incorporation of the precursor
monomer subunit into the strand, a ligand having an electrophilic
group, e.g., a pentafluorophenyl ester or aldehyde group, can
subsequently be attached to the precursor ligand-conjugated monomer
by coupling the electrophilic group of the ligand with the terminal
nucleophilic group of the precursor ligand-conjugated monomer
subunit tether.
[0113] In preferred embodiments, a ligand alters the distribution,
targeting or lifetime of an iRNA agent into which it is
incorporated. In preferred embodiments a ligand provides an
enhanced affinity for a selected target, e.g., molecule, cell or
cell type, compartment, e.g., a cellular or organ compartment,
tissue, organ or region of the body, as, e.g., compared to a
species absent such a ligand.
[0114] Preferred ligands can improve transport, hybridization, and
specificity properties and may also improve nuclease resistance of
the resultant natural or modified oligoribonucleotide, or a
polymeric molecule comprising any combination of monomers described
herein and/or natural or modified ribonucleotides.
[0115] Ligands in general can include therapeutic modifiers, e.g.,
for enhancing uptake; diagnostic compounds or reporter groups e.g.,
for monitoring distribution; cross-linking agents;
nuclease-resistance conferring moieties; and natural or unusual
nucleobases. General examples include lipophilic molecules, lipids,
lectins, steroids (e.g., uvaol, hecigenin, diosgenin), terpenes
(e.g., triterpenes, e.g., sarsasapogenin, Friedelin, epifriedelanol
derivatized lithocholic acid), vitamins, carbohydrates (e.g., a
dextran, pullulan, chitin, chitosan, inulin, cyclodextrin or
hyaluronic acid), proteins, protein binding agents, integrin
targeting molecules, polycationics, peptides, polyamines, and
peptide mimics.
[0116] The ligand may be a naturally occurring or recombinant or
synthetic molecule, such as a synthetic polymer, e.g., a synthetic
polyamino acid. Examples of polyamino acids include polylysine
(PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic
acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer,
divinyl ether-maleic anhydride copolymer,
N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene
glycol (PEG), polyvinyl alcohol (PVA), polyurethane,
poly(2-ethylacrylic acid), N-isopropylacrylamide polymers, or
polyphosphazine. Example of polyamines include: polyethylenimine,
polylysine (PLL), spermine, spermidine, polyamine,
pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer
polyamine, arginine, amidine, protamine, cationic moieties, e.g.,
cationic lipid, cationic porphyrin, quaternary salt of a polyamine,
or an alpha helical peptide.
[0117] Ligands can also include targeting groups, e.g., a cell or
tissue targeting agent, e.g., a thyrotropin, melanotropin,
surfactant protein A, Mucin carbohydrate, a glycosylated
polyaminoacid, transferrin, bisphosphonate, polyglutamate,
polyaspartate, or an RGD peptide or RGD peptide mimetic.
[0118] Ligands can be proteins, e.g., glycoproteins, lipoproteins,
e.g. low density lipoprotein (LDL), or albumins, e.g. human serum
albumin (HSA), or peptides, e.g., molecules having a specific
affinity for a co-ligand, or antibodies e.g., an antibody, that
binds to a specified cell type such as a cancer cell, endothelial
cell, or bone cell. Ligands may also include hormones and hormone
receptors. They can also include non-peptidic species, such as
cofactors, multivalent lactose, multivalent galactose,
N-acetyl-galactosamine, N-acetyl-glucosamine, multivalent mannose,
or multivalent fucose. The ligand can be, for example, a
lipopolysaccharide, an activator of p38 MAP kinase, or an activator
of NF-.kappa.B.
[0119] The ligand can be a substance, e.g, a drug, which can
increase the uptake of the iRNA agent into the cell, for example,
by disrupting the cell's cytoskeleton, e.g., by disrupting the
cell's microtubules, microfilaments, and/or intermediate filaments.
The drug can be, for example, taxon, vincristine, vinblastine,
cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin,
swinholide A, indanocine, or myoservin.
[0120] In one aspect, the ligand is a lipid or lipid-based
molecule. Such a lipid or lipid-based molecule preferably binds a
serum protein, e.g., human serum albumin (HSA). An HSA binding
ligand allows for distribution of the conjugate to a target tissue,
e.g., liver tissue, including parenchymal cells of the liver. Other
molecules that can bind HSA can also be used as ligands. For
example, neproxin or aspirin can be used. A lipid or lipid-based
ligand can (a) increase resistance to degradation of the conjugate,
(b) increase targeting or transport into a target cell or cell
membrane, and/or (c) can be used to adjust binding to a serum
protein, e.g., HSA.
[0121] A lipid based ligand can be used to modulate, e.g., control
the binding of the conjugate to a target tissue. For example, a
lipid or lipid-based ligand that binds to HSA more strongly will be
less likely to be targeted to the kidney and therefore less likely
to be cleared from the body. A lipid or lipid-based ligand that
binds to HSA less strongly can be used to target the conjugate to
the kidney.
[0122] In a preferred embodiment, the lipid based ligand binds HSA.
Preferably, it binds HSA with a sufficient affinity such that the
conjugate will be preferably distributed to a non-kidney tissue.
However, it is preferred that the affinity not be so strong that
the HSA-ligand binding cannot be reversed.
[0123] In another aspect, the ligand is a moiety, e.g., a vitamin
or nutrient, which is taken up by a target cell, e.g., a
proliferating cell. These are particularly useful for treating
disorders characterized by unwanted cell proliferation, e.g., of
the malignant or non-malignant type, e.g., cancer cells. Exemplary
vitamins include vitamin A, E, and K. Other exemplary vitamins
include the B vitamins, e.g., folic acid, B12, riboflavin, biotin,
pyridoxal or other vitamins or nutrients taken up by cancer
cells.
[0124] In another aspect, the ligand is a cell-permeation agent,
preferably a helical cell-permeation agent. Preferably, the agent
is amphipathic. An exemplary agent is a peptide such as tat or
antennapedia. If the agent is a peptide, it can be modified,
including a peptidylmimetic, invertomers, non-peptide or
pseudo-peptide linkages, and use of D-amino acids. The helical
agent is preferably an alpha-helical agent, which preferably has a
lipophilic and a lipophobic phase.
[0125] 5'-Phosphate Modifications
[0126] In preferred embodiments, iRNA agents are 5' phosphorylated
or include a phosphoryl analog at the 5' prime terminus.
5'-phosphate modifications of the antisense strand include those
which are compatible with RISC mediated gene silencing. Suitable
modifications include: 5'-monophosphate ((HO)2(O)P--O-5');
5'-diphosphate ((HO)2(O)P--O--P(HO)(O)--O-5'); 5'-triphosphate
((HO)2(O)P--O--(HO)(O)P--O--P(HO)(O)--O-5'); 5'-guanosine cap
(7-methylated or non-methylated)
(7m-G-O-5'-(HO)(O)P--O--(HO)(O)P--O--P(HO)(O)--O-5'); 5'-adenosine
cap (Appp), and any modified or unmodified nucleotide cap structure
(N--O-5'-(HO)(O)P--O--(HO)(O)P--O--P(HO)(O)--O-5');
5'-monothiophosphate (phosphorothioate; (HO)2(S)P--O-5');
5'-monodithiophosphate (phosphorodithioate; (HO)(HS)(S)P--O-5'),
5'-phosphorothiolate ((HO)2(O)P--S-5'); any additional combination
of oxygen/sulfur replaced monophosphate, diphosphate and
triphosphates (e.g. 5'-alpha-thiotriphosphate,
5'-gamma-thiotriphosphate, etc.), 5'-phosphoramidates
((HO)2(O)P--NH-5', (HO)(NH2)(O)P--O-5'), 5'-alkylphosphonates
(R=alkyl=methyl, ethyl, isopropyl, propyl, etc., e.g.
RP(OH)(O)--O-5'-, (OH)2(O)P-5'-CH2-), 5'-alkyletherphosphonates
(R=alkylether=methoxymethyl (MeOCH2-), ethoxymethyl, etc., e.g.
RP(OH)(O)--O-5'-).
[0127] The sense strand can be modified in order to inactivate the
sense strand and prevent formation of an active RISC, thereby
potentially reducing off-target effects. This can be accomplished
by a modification which prevents 5'-phosphorylation of the sense
strand, e.g., by modification with a 5'-O-methyl ribonucleotide
(see Nykanen et al., (2001) ATP requirements and small interfering
RNA structure in the RNA interference pathway. Cell 107, 309-321.)
Other modifications which prevent phosphorylation can also be used,
e.g., simply substituting the 5'-OH by H rather than O-Me.
Alternatively, a large bulky group may be added to the 5'-phosphate
turning it into a phosphodiester linkage.
[0128] Non-Natural Nucleobases
[0129] Nitropyrrolyl and nitroindolyl are non-natural nucleobases
that are members of a class of compounds known as universal bases.
Universal bases are those compounds that can replace any of the
four naturally occurring bases without substantially affecting the
melting behavior or activity of the oligonucleotide duplex. In
contrast to the stabilizing, hydrogen-bonding interactions
associated with naturally occurring nucleobases, it is postulated
that oligonucleotide duplexes containing 3-nitropyrrolyl
nucleobases are stabilized solely by stacking interactions. The
absence of significant hydrogen-bonding interactions with
nitropyrrolyl nucleobases obviates the specificity for a specific
complementary base. In addition, various reports confirm that 4-,
5- and 6-nitroindolyl display very little specificity for the four
natural bases. Interestingly, an oligonucleotide duplex containing
5-nitroindolyl was more stable than the corresponding
oligonucleotides containing 4-nitroindolyl and 6-nitroindolyl.
Procedures for the preparation of
1-(2'-O-methyl-.beta.-D-ribofuranosyl)-5-nitroindole are described
in Gaubert, G.; Wengel, J. Tetrahedron Letters 2004, 45, 5629.
Other universal bases amenable to the present invention include
hypoxanthinyl, isoinosinyl, 2-aza-inosinyl, 7-deaza-inosinyl,
nitroimidazolyl, nitropyrazolyl, nitrobenzimidazolyl,
nitroindazolyl, aminoindolyl, pyrrolopyrimidinyl, and structural
derivatives thereof. For a more detailed discussion, including
synthetic procedures, of nitropyrrolyl, nitroindolyl, and other
universal bases mentioned above see Vallone et al., Nucleic Acids
Research, 27(17):3589-3596 (1999); Loakes et al., J. Mol. Bio.,
270:426-436 (1997); Loakes et al., Nucleic Acids Research,
22(20):4039-4043 (1994); Oliver et al., Organic Letters, Vol.
3(13):1977-1980 (2001); Amosova et al., Nucleic Acids Research,
25(10):1930-1934 (1997); Loakes et al., Nucleic Acids Research,
29(12):2437-2447 (2001); Bergstrom et al., J. Am. Chem. Soc.,
117:1201-1209 (1995); Franchetti et al., Biorg. Med. Chem. Lett.
11:67-69 (2001); and Nair et al., Nucelosides, Nucleotides &
Nucleic Acids, 20(4-7):735-738 (2001).
[0130] Difluorotolyl is a non-natural nucleobase that functions as
a universal base. Difluorotolyl is an isostere of the natural
nucleobase thymine. But unlike thymine, difluorotolyl shows no
appreciable selectivity for any of the natural bases. Other
aromatic compounds that function as universal bases and are
amenable to the present invention are
4-fluoro-6-methylbenzimidazole and 4-methylbenzimidazole. In
addition, the relatively hydrophobic isocarbostyrilyl derivatives
3-methyl isocarbostyrilyl, 5-methyl isocarbostyrilyl, and
3-methyl-7-propynyl isocarbostyrilyl are universal bases which
cause only slight destabilization of oligonucleotide duplexes
compared to the oligonucleotide sequence containing only natural
bases. Other non-natural nucleobases contemplated in the present
invention include 7-azaindolyl, 6-methyl-7-azaindolyl,
imidizopyridinyl, 9-methyl-imidizopyridinyl, pyrrolopyrizinyl,
isocarbostyrilyl, 7-propynyl isocarbostyrilyl,
propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylindolyl,
4,6-dimethylindolyl, phenyl, napthalenyl, anthracenyl,
phenanthracenyl, pyrenyl, stilbenyl, tetracenyl, pentacenyl, and
structural derivates thereof. For a more detailed discussion,
including synthetic procedures, of difluorotolyl,
4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, and other
non-natural bases mentioned above, see: Schweitzer et al., J. Org.
Chem., 59:7238-7242 (1994); Berger et al., Nucleic Acids Research,
28(15):2911-2914 (2000); Moran et al., J. Am. Chem. Soc.,
119:2056-2057 (1997); Morales et al., J. Am. Chem. Soc.,
121:2323-2324 (1999); Guckian et al., J. Am. Chem. Soc.,
118:8182-8183 (1996); Morales et al., J. Am. Chem. Soc.,
122(6):1001-1007 (2000); McMinn et al., J. Am. Chem. Soc.,
121:11585-11586 (1999); Guckian et al., J. Org. Chem., 63:9652-9656
(1998); Moran et al., Proc. Natl. Acad. Sci., 94:10506-10511
(1997); Das et al., J. Chem. Soc., Perkin Trans., 1:197-206 (2002);
Shibata et al., J. Chem. Soc., Perkin Trans., 1: 1605-1611 (2001);
Wu et al., J. Am. Chem. Soc., 122(32):7621-7632 (2000); O'Neill et
al., J. Org. Chem., 67:5869-5875 (2002); Chaudhuri et al., J. Am.
Chem. Soc., 117:10434-10442 (1995); and U.S. Pat. No.
6,218,108.
[0131] Further details to the synthesis and use of universal bases
is given in co-owned and co-pending PCT/US2005/025967, filed Jul.
21, 2005, hereby incorporated herein by reference in its
entirety.
[0132] Universal bases can be particularly helpful in situations
where one attempts to target a gene in an organism that shows a
variability between different strains of that organism. This is
often true even for regions of a viral genome that are regarded as
highly conserved. The incorporation of a universal base may allow
the design of iRNA agents that target a large number of different
strains of a virus even though they differ in one, or a few, e.g.
in up to three, nucleotide positions.
[0133] Universal bases are best included in a region of an iRNA
agent that is least sensitive to nucleotide mismatches with regard
to specifity and activity of the iRNA agent. It has been shown that
position 2-9 of the antisense strand of an iRNA agent are most
sensitive to mismatches between the antisense strand an the target
mRNA, and this region has been termed the "seed-region" of an iRNA
agent. Hence, when incorporating one or several universal base or
bases into an iRNA agent, it or they are preferably incorporated
outside this seed region.
[0134] Table 1F and Table 1G show iRNA agents comprising universal
bases in mutually complementary positions in the sense and
antisense strand. However, while this is one preferred embodiment
of the iRNA agents of the present invention, the effect of the
universal base in an iRNA agent is more pronounced when the
universal base is present in the antisense strand. It is therefore
envisioned that the base in the sense strand in a position where it
will pair up with a universal base in the antisense strand may be
either a universal base, or any other suitable base, such as a, u,
c or g. Preferably, one will test which base in such position of
the sense strand will give the highest activity and/or selectivity
for the iRNA agent. Alternatively, the base may be chosen that is
present in this particular position in a majority of the target
gene variants intended to be inhibited in their expression by the
iRNA agent in question.
[0135] Transport of iRNA Agents into Cells
[0136] Not wishing to be bound by any theory, the chemical
similarity between cholesterol-conjugated iRNA agents and certain
constituents of lipoproteins (e.g. cholesterol, cholesteryl esters,
phospholipids) may lead to the association of iRNA agents with
lipoproteins (e.g. LDL, HDL) in blood and/or the interaction of the
iRNA agent with cellular components having an affinity for
cholesterol, e.g. components of the cholesterol transport pathway.
Lipoproteins as well as their constituents are taken up and
processed by cells by various active and passive transport
mechanisms, for example, without limitation, endocytosis of
LDL-receptor bound LDL, endocytosis of oxidized or otherwise
modified LDLs through interaction with Scavenger receptor A,
Scavenger receptor B1-mediated uptake of HDL cholesterol in the
liver, pinocytosis, or transport of cholesterol across membranes by
ABC (ATP-binding cassette) transporter proteins, e.g. ABC-A1,
ABC-G1 or ABC-G4. Hence, cholesterol-conjugated iRNA agents could
enjoy facilitated uptake by cells possessing such transport
mechanisms, e.g. cells of the liver. As such, the present invention
provides evidence and general methods for targeting iRNA agents to
cells expressing certain cell surface components, e.g. receptors,
by conjugating a natural ligand for such component (e.g.
cholesterol) to the iRNA agent, or by conjugating a chemical moiety
(e.g. cholesterol) to the iRNA agent which associates with or binds
to a natural ligand for the component (e.g. LDL, HDL).
Other Embodiments
[0137] An RNA, e.g., an iRNA agent, can be produced in a cell in
vivo, e.g., from exogenous DNA templates that are delivered into
the cell. For example, the DNA templates can be inserted into
vectors and used as gene therapy vectors. Gene therapy vectors can
be delivered to a subject by, for example, intravenous injection,
local administration (U.S. Pat. No. 5,328,470), or by stereotactic
injection (see, e.g., Chen et al. Proc. Natl. Acad. Sci. USA
91:3054-3057, 1994). The pharmaceutical preparation of the gene
therapy vector can include the gene therapy vector in an acceptable
diluent, or can comprise a slow release matrix in which the gene
delivery vehicle is imbedded. The DNA templates, for example, can
include two transcription units, one that produces a transcript
that includes the top strand of an iRNA agent and one that produces
a transcript that includes the bottom strand of an iRNA agent. When
the templates are transcribed, the iRNA agent is produced, and
processed into siRNA agent fragments that mediate gene
silencing.
[0138] Formulation
[0139] The present invention also includes pharmaceutical
compositions and formulations which include the dsRNA compounds of
the invention. The pharmaceutical compositions of the present
invention may be administered in a number of ways depending upon
whether local or systemic treatment is desired and upon the area to
be treated. Administration may be topical, pulmonary, e.g., by
inhalation or insufflation of powders or aerosols, including by
nebulizer; intratracheal, intranasal, epidermal and transdermal,
oral or parenteral. Parenteral administration includes intravenous,
intraarterial, subcutaneous, intraperitoneal or intramuscular
injection or infusion; or intracranial, e.g., intrathecal or
intraventricular, administration.
[0140] Pharmaceutical compositions and formulations for topical
administration may include transdermal patches, ointments, lotions,
creams, gels, drops, suppositories, sprays, liquids and powders.
Conventional pharmaceutical carriers, aqueous, powder or oily
bases, thickeners and the like may be necessary or desirable.
Coated condoms, gloves and the like may also be useful. Preferred
topical formulations include those in which the dsRNAs of the
invention are in admixture with a topical delivery agent such as
lipids, liposomes, fatty acids, fatty acid esters, steroids,
chelating agents and surfactants. Preferred lipids and liposomes
include neutral (e.g. dioleoylphosphatidyl ethanolamine=DOPE,
dimyristoylphosphatidyl choline=DMPC, distearolyphosphatidyl
choline) negative (e.g. dimyristoylphosphatidyl glycerol=DMPG) and
cationic (e.g. dioleoyltetramethylaminopropyl=DOTAP and
dioleoylphosphatidyl ethanolamine=DOTMA). DsRNAs of the invention
may be encapsulated within liposomes or may form complexes thereto,
in particular to cationic liposomes. Alternatively, dsRNAs may be
complexed to lipids, in particular to cationic lipids. Preferred
fatty acids and esters include but are not limited arachidonic
acid, oleic acid, eicosanoic acid, lauric acid, caprylic acid,
capric acid, myristic acid, palmitic acid, stearic acid, linoleic
acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin,
glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an
acylcarnitine, an acylcholine, or a C.sub.1-10 alkyl ester (e.g.
isopropylmyristate IPM), monoglyceride, diglyceride or
pharmaceutically acceptable salt thereof. Topical formulations are
described in detail in U.S. patent application Ser. No. 09/315,298
filed on May 20, 1999 which is incorporated herein by reference in
its entirety.
[0141] Compositions and formulations for oral administration
include powders or granules, microparticulates, nanoparticulates,
suspensions or solutions in water or non-aqueous media, capsules,
gel capsules, sachets, tablets or minitablets. Thickeners,
flavoring agents, diluents, emulsifiers, dispersing aids or binders
may be desirable. Preferred oral formulations are those in which
dsRNAs of the invention are administered in conjunction with one or
more penetration enhancers, surfactants, and chelators. Preferred
surfactants include fatty acids and/or esters or salts thereof,
bile acids and/or salts thereof. Preferred bile acids/salts include
chenodeoxycholic acid (CDCA) and ursodeoxychenodeoxycholic acid
(UDCA), cholic acid, dehydrocholic acid, deoxycholic acid,
glucholic acid, glycholic acid, glycodeoxycholic acid, taurocholic
acid, taurodeoxycholic acid, sodium tauro-24,25-dihydro-fusidate
and sodium glycodihydrofusidate. Preferred fatty acids include
arachidonic acid, undecanoic acid, oleic acid, lauric acid,
caprylic acid, capric acid, myristic acid, palmitic acid, stearic
acid, linoleic acid, linolenic acid, dicaprate, tricaprate,
monoolein, dilaurin, glyceryl 1-monocaprate,
1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or
a monoglyceride, a diglyceride or a pharmaceutically acceptable
salt thereof (e.g. sodium). Also preferred are combinations of
penetration enhancers, for example, fatty acids/salts in
combination with bile acids/salts. A particularly preferred
combination is the sodium salt of lauric acid, capric acid and
UDCA. Further penetration enhancers include
polyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether.
DsRNAs of the invention may be delivered orally, in granular form
including sprayed dried particles, or complexed to form micro or
nanoparticles. DsRNA complexing agents include poly-amino acids;
polyimines; polyacrylates; polyalkylacrylates, polyoxethanes,
polyalkylcyanoacrylates; cationized gelatins, albumins, starches,
acrylates, polyethyleneglycols (PEG) and starches;
polyalkylcyanoacrylates; DEAE-derivatized polyimines, pollulans,
celluloses and starches. Particularly preferred complexing agents
include chitosan, N-trimethylchitosan, poly-L-lysine,
polyhistidine, polyornithine, polyspermines, protamine,
polyvinylpyridine, polythiodiethylaminomethylethylene P(TDAE),
polyaminostyrene (e.g. p-amino), poly(methylcyanoacrylate),
poly(ethylcyanoacrylate), poly(butylcyanoacrylate),
poly(isobutylcyanoacrylate), poly(isohexylcynaoacrylate),
DEAE-methacrylate, DEAE-hexylacrylate, DEAE-acrylamide,
DEAE-albumin and DEAE-dextran, polymethylacrylate,
polyhexylacrylate, poly(D,L-lactic acid),
poly(DL-lactic-co-glycolic acid (PLGA), alginate, and
polyethyleneglycol (PEG). Oral formulations for dsRNAs and their
preparation are described in detail in U.S. application. Ser. No.
08/886,829 (filed Jul. 1, 1997), Ser. No. 09/108,673 (filed Jul. 1,
1998), Ser. No. 09/256,515 (filed Feb. 23, 1999), Ser. No.
09/082,624 (filed May 21, 1998) and Ser. No. 09/315,298 (filed May
20, 1999), each of which is incorporated herein by reference in
their entirety.
[0142] Compositions and formulations for parenteral, intrathecal or
intraventricular administration may include sterile aqueous
solutions which may also contain buffers, diluents and other
suitable additives such as, but not limited to, penetration
enhancers, carrier compounds and other pharmaceutically acceptable
carriers or excipients.
[0143] Pharmaceutical compositions of the present invention
include, but are not limited to, solutions, emulsions, and
liposome-containing formulations. These compositions may be
generated from a variety of components that include, but are not
limited to, preformed liquids, self-emulsifying solids and
self-emulsifying semisolids.
[0144] The pharmaceutical formulations of the present invention,
which may conveniently be presented in unit dosage form, may be
prepared according to conventional techniques well known in the
pharmaceutical industry. Such techniques include the step of
bringing into association the active ingredients with the
pharmaceutical carrier(s) or excipient(s). In general, the
formulations are prepared by uniformly and intimately bringing into
association the active ingredients with liquid carriers or finely
divided solid carriers or both, and then, if necessary, shaping the
product.
[0145] The compositions of the present invention may be formulated
into any of many possible dosage forms such as, but not limited to,
tablets, capsules, gel capsules, liquid syrups, soft gels,
suppositories, and enemas. The compositions of the present
invention may also be formulated as suspensions in aqueous,
non-aqueous or mixed media. Aqueous suspensions may further contain
substances which increase the viscosity of the suspension
including, for example, sodium carboxymethylcellulose, sorbitol
and/or dextran. The suspension may also contain stabilizers.
[0146] In one embodiment of the present invention the
pharmaceutical compositions may be formulated and used as foams.
Pharmaceutical foams include formulations such as, but not limited
to, emulsions, microemulsions, creams, jellies and liposomes. While
basically similar in nature these formulations vary in the
components and the consistency of the final product. The
preparation of such compositions and formulations is generally
known to those skilled in the pharmaceutical and formulation arts
and may be applied to the formulation of the compositions of the
present invention.
[0147] Emulsions
[0148] The compositions of the present invention may be prepared
and formulated as emulsions. Emulsions are typically heterogenous
systems of one liquid dispersed in another in the form of droplets
usually exceeding 0.1 .mu.m in diameter (Idson, in Pharmaceutical
Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel
Dekker, Inc., New York, N.Y., volume 1, p. 199; Rosoff, in
Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.),
1988, Marcel Dekker, Inc., New York, N.Y., Volume 1, p. 245; Block
in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker
(Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 2, p.
335; Higuchi et al., in Remington's Pharmaceutical Sciences, Mack
Publishing Co., Easton, Pa., 1985, p. 301). Emulsions are often
biphasic systems comprising two immiscible liquid phases intimately
mixed and dispersed with each other. In general, emulsions may be
of either the water-in-oil (w/o) or the oil-in-water (o/w) variety.
When an aqueous phase is finely divided into and dispersed as
minute droplets into a bulk oily phase, the resulting composition
is called a water-in-oil (w/o) emulsion. Alternatively, when an
oily phase is finely divided into and dispersed as minute droplets
into a bulk aqueous phase, the resulting composition is called an
oil-in-water (o/w) emulsion. Emulsions may contain additional
components in addition to the dispersed phases, and the active drug
which may be present as a solution in either the aqueous phase,
oily phase or itself as a separate phase. Pharmaceutical excipients
such as emulsifiers, stabilizers, dyes, and anti-oxidants may also
be present in emulsions as needed. Pharmaceutical emulsions may
also be multiple emulsions that are comprised of more than two
phases such as, for example, in the case of oil-in-water-in-oil
(o/w/o) and water-in-oil-in-water (w/o/w) emulsions. Such complex
formulations often provide certain advantages that simple binary
emulsions do not. Multiple emulsions in which individual oil
droplets of an o/w emulsion enclose small water droplets constitute
a w/o/w emulsion. Likewise a system of oil droplets enclosed in
globules of water stabilized in an oily continuous phase provides
an o/w/o emulsion.
[0149] Emulsions are characterized by little or no thermodynamic
stability. Often, the dispersed or discontinuous phase of the
emulsion is well dispersed into the external or continuous phase
and maintained in this form through the means of emulsifiers or the
viscosity of the formulation. Either of the phases of the emulsion
may be a semisolid or a solid, as is the case of emulsion-style
ointment bases and creams. Other means of stabilizing emulsions
entail the use of emulsifiers that may be incorporated into either
phase of the emulsion. Emulsifiers may broadly be classified into
four categories: synthetic surfactants, naturally occurring
emulsifiers, absorption bases, and finely dispersed solids (Idson,
in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker
(Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.
199).
[0150] Synthetic surfactants, also known as surface active agents,
have found wide applicability in the formulation of emulsions and
have been reviewed in the literature (Rieger, in Pharmaceutical
Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel
Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in
Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.),
Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199).
Surfactants are typically amphiphilic and comprise a hydrophilic
and a hydrophobic portion. The ratio of the hydrophilic to the
hydrophobic nature of the surfactant has been termed the
hydrophile/lipophile balance (HLB) and is a valuable tool in
categorizing and selecting surfactants in the preparation of
formulations. Surfactants may be classified into different classes
based on the nature of the hydrophilic group: nonionic, anionic,
cationic and amphoteric (Rieger, in Pharmaceutical Dosage Forms,
Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New
York, N.Y., volume 1, p. 285).
[0151] Naturally occurring emulsifiers used in emulsion
formulations include lanolin, beeswax, phosphatides, lecithin and
acacia. Absorption bases possess hydrophilic properties such that
they can soak up water to form w/o emulsions yet retain their
semisolid consistencies, such as anhydrous lanolin and hydrophilic
petrolatum. Finely divided solids have also been used as good
emulsifiers especially in combination with surfactants and in
viscous preparations. These include polar inorganic solids, such as
heavy metal hydroxides, nonswelling clays such as bentonite,
attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum
silicate and colloidal magnesium aluminum silicate, pigments and
nonpolar solids such as carbon or glyceryl tristearate.
[0152] A large variety of non-emulsifying materials are also
included in emulsion formulations and contribute to the properties
of emulsions. These include fats, oils, waxes, fatty acids, fatty
alcohols, fatty esters, humectants, hydrophilic colloids,
preservatives and antioxidants (Block, in Pharmaceutical Dosage
Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker,
Inc., New York, N.Y., volume 1, p. 335; Idson, in Pharmaceutical
Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel
Dekker, Inc., New York, N.Y., volume 1, p. 199).
[0153] Hydrophilic colloids or hydrocolloids include naturally
occurring gums and synthetic polymers such as polysaccharides (for
example, acacia, agar, alginic acid, carrageenan, guar gum, karaya
gum, and tragacanth), cellulose derivatives (for example,
carboxymethylcellulose and carboxypropylcellulose), and synthetic
polymers (for example, carbomers, cellulose ethers, and
carboxyvinyl polymers). These disperse or swell in water to form
colloidal solutions that stabilize emulsions by forming strong
interfacial films around the dispersed-phase droplets and by
increasing the viscosity of the external phase.
[0154] Since emulsions often contain a number of ingredients such
as carbohydrates, proteins, sterols and phosphatides that may
readily support the growth of microbes, these formulations often
incorporate preservatives. Commonly used preservatives included in
emulsion formulations include methyl paraben, propyl paraben,
quaternary ammonium salts, benzalkonium chloride, esters of
p-hydroxybenzoic acid, and boric acid. Antioxidants are also
commonly added to emulsion formulations to prevent deterioration of
the formulation. Antioxidants used may be free radical scavengers
such as tocopherols, alkyl gallates, butylated hydroxyanisole,
butylated hydroxytoluene, or reducing agents such as ascorbic acid
and sodium metabisulfite, and antioxidant synergists such as citric
acid, tartaric acid, and lecithin.
[0155] The application of emulsion formulations via dermatological,
oral and parenteral routes and methods for their manufacture have
been reviewed in the literature (Idson, in Pharmaceutical Dosage
Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker,
Inc., New York, N.Y., volume 1, p. 199). Emulsion formulations for
oral delivery have been very widely used because of ease of
formulation, as well as efficacy from an absorption and
bioavailability standpoint (Rosoff, in Pharmaceutical Dosage Forms,
Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New
York, N.Y., volume 1, p. 245; Idson, in Pharmaceutical Dosage
Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker,
Inc., New York, N.Y., volume 1, p. 199). Mineral-oil base
laxatives, oil-soluble vitamins and high fat nutritive preparations
are among the materials that have commonly been administered orally
as o/w emulsions.
[0156] In one embodiment of the present invention, the compositions
of dsRNAs and nucleic acids are formulated as microemulsions. A
microemulsion may be defined as a system of water, oil and
amphiphile which is a single optically isotropic and
thermodynamically stable liquid solution (Rosoff, in Pharmaceutical
Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel
Dekker, Inc., New York, N.Y., volume 1, p. 245). Typically
microemulsions are systems that are prepared by first dispersing an
oil in an aqueous surfactant solution and then adding a sufficient
amount of a fourth component, generally an intermediate
chain-length alcohol to form a transparent system. Therefore,
microemulsions have also been described as thermodynamically
stable, isotropically clear dispersions of two immiscible liquids
that are stabilized by interfacial films of surface-active
molecules (Leung and Shah, in: Controlled Release of Drugs:
Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH
Publishers, New York, pages 185-215). Microemulsions commonly are
prepared via a combination of three to five components that include
oil, water, surfactant, cosurfactant and electrolyte. Whether the
microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w)
type is dependent on the properties of the oil and surfactant used
and on the structure and geometric packing of the polar heads and
hydrocarbon tails of the surfactant molecules (Schott, in
Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,
Pa., 1985, p. 271).
[0157] The phenomenological approach utilizing phase diagrams has
been extensively studied and has yielded a comprehensive knowledge,
to one skilled in the art, of how to formulate microemulsions
(Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and
Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1,
p. 245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger
and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y.,
volume 1, p. 335). Compared to conventional emulsions,
microemulsions offer the advantage of solubilizing water-insoluble
drugs in a formulation of thermodynamically stable droplets that
are formed spontaneously.
[0158] Surfactants used in the preparation of microemulsions
include, but are not limited to, ionic surfactants, non-ionic
surfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol
fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol
monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol
pentaoleate (PO500), decaglycerol monocaprate (MCA750),
decaglycerol monooleate (MO750), decaglycerol sequioleate (SO750),
decaglycerol decaoleate (DA0750), alone or in combination with
cosurfactants. The cosurfactant, usually a short-chain alcohol such
as ethanol, 1-propanol, and 1-butanol, serves to increase the
interfacial fluidity by penetrating into the surfactant film and
consequently creating a disordered film because of the void space
generated among surfactant molecules. Microemulsions may, however,
be prepared without the use of cosurfactants and alcohol-free
self-emulsifying microemulsion systems are known in the art. The
aqueous phase may typically be, but is not limited to, water, an
aqueous solution of the drug, glycerol, PEG300, PEG400,
polyglycerols, propylene glycols, and derivatives of ethylene
glycol. The oil phase may include, but is not limited to, materials
such as Captex 300, Captex 355, Capmul MCM, fatty acid esters,
medium chain (C.sub.8-C.sub.12) mono, di, and tri-glycerides,
polyoxyethylated glyceryl fatty acid esters, fatty alcohols,
polyglycolized glycerides, saturated polyglycolized
C.sub.8-C.sub.10 glycerides, vegetable oils and silicone oil.
[0159] Microemulsions are particularly of interest from the
standpoint of drug solubilization and the enhanced absorption of
drugs. Lipid based microemulsions (both o/w and w/o) have been
proposed to enhance the oral bioavailability of drugs, including
peptides (Constantinides et al., Pharmaceutical Research, 1994, 11,
1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol., 1993, 13,
205). Microemulsions afford advantages of improved drug
solubilization, protection of drug from enzymatic hydrolysis,
possible enhancement of drug absorption due to surfactant-induced
alterations in membrane fluidity and permeability, ease of
preparation, ease of oral administration over solid dosage forms,
improved clinical potency, and decreased toxicity (Constantinides
et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J.
Pharm. Sci., 1996, 85, 138-143). Often microemulsions may form
spontaneously when their components are brought together at ambient
temperature. This may be particularly advantageous when formulating
thermolabile drugs, peptides or dsRNAs. Microemulsions have also
been effective in the transdermal delivery of active components in
both cosmetic and pharmaceutical applications. It is expected that
the microemulsion compositions and formulations of the present
invention will facilitate the increased systemic absorption of
dsRNAs and nucleic acids from the gastrointestinal tract, as well
as improve the local cellular uptake of dsRNAs and nucleic acids
within the gastrointestinal tract, vagina, buccal cavity and other
areas of administration.
[0160] Microemulsions of the present invention may also contain
additional components and additives such as sorbitan monostearate
(Grill 3), Labrasol, and penetration enhancers to improve the
properties of the formulation and to enhance the absorption of the
dsRNAs and nucleic acids of the present invention. Penetration
enhancers used in the microemulsions of the present invention may
be classified as belonging to one of five broad categories
surfactants, fatty acids, bile salts, chelating agents, and
non-chelating non-surfactants (Lee et al., Critical Reviews in
Therapeutic Drug Carrier Systems, 1991, p. 92). Each of these
classes has been discussed above.
[0161] Liposomes
[0162] There are many organized surfactant structures besides
microemulsions that have been studied and used for the formulation
of drugs. These include monolayers, micelles, bilayers and
vesicles. Vesicles, such as liposomes, have attracted great
interest because of their specificity and the duration of action
they offer from the standpoint of drug delivery. As used in the
present invention, the term "liposome" means a vesicle composed of
amphiphilic lipids arranged in a spherical bilayer or bilayers.
[0163] Liposomes are unilamellar or multilamellar vesicles which
have a membrane formed from a lipophilic material and an aqueous
interior. The aqueous portion contains the composition to be
delivered. Cationic liposomes possess the advantage of being able
to fuse to the cell wall. Non-cationic liposomes, although not able
to fuse as efficiently with the cell wall, are taken up by
macrophages in vivo.
[0164] In order to cross intact mammalian skin, lipid vesicles must
pass through a series of fine pores, each with a diameter less than
50 nm, under the influence of a suitable transdermal gradient.
Therefore, it is desirable to use a liposome which is highly
deformable and able to pass through such fine pores.
[0165] Further advantages of liposomes include; liposomes obtained
from natural phospholipids are biocompatible and biodegradable;
liposomes can incorporate a wide range of water and lipid soluble
drugs; liposomes can protect encapsulated drugs in their internal
compartments from metabolism and degradation (Rosoff, in
Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.),
1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245).
Important considerations in the preparation of liposome
formulations are the lipid surface charge, vesicle size and the
aqueous volume of the liposomes.
[0166] Liposomes are useful for the transfer and delivery of active
ingredients to the site of action. Because the liposomal membrane
is structurally similar to biological membranes, when liposomes are
applied to a tissue, the liposomes start to merge with the cellular
membranes and as the merging of the liposome and cell progresses,
the liposomal contents are emptied into the cell where the active
agent may act.
[0167] Liposomal formulations have been the focus of extensive
investigation as the mode of delivery for many drugs. There is
growing evidence that for topical administration, liposomes present
several advantages over other formulations. Such advantages include
reduced side-effects related to high systemic absorption of the
administered drug, increased accumulation of the administered drug
at the desired target, and the ability to administer a wide variety
of drugs, both hydrophilic and hydrophobic, into the skin.
[0168] Several reports have detailed the ability of liposomes to
deliver agents including high-molecular weight DNA into the skin.
Compounds including analgesics, antibodies, hormones and
high-molecular weight DNAs have been administered to the skin. The
majority of applications resulted in the targeting of the upper
epidermis
[0169] Liposomes fall into two broad classes. Cationic liposomes
are positively charged liposomes which interact with the negatively
charged DNA molecules to form a stable complex. The positively
charged DNA/liposome complex binds to the negatively charged cell
surface and is internalized in an endosome. Due to the acidic pH
within the endosome, the liposomes are ruptured, releasing their
contents into the cell cytoplasm (Wang et al., Biochem. Biophys.
Res. Commun., 1987, 147, 980-985).
[0170] Liposomes which are pH-sensitive or negatively-charged,
entrap DNA rather than complex with it. Since both the DNA and the
lipid are similarly charged, repulsion rather than complex
formation occurs. Nevertheless, some DNA is entrapped within the
aqueous interior of these liposomes. pH-sensitive liposomes have
been used to deliver DNA encoding the thymidine kinase gene to cell
monolayers in culture. Expression of the exogenous gene was
detected in the target cells (Zhou et al., Journal of Controlled
Release, 1992, 19, 269-274).
[0171] One major type of liposomal composition includes
phospholipids other than naturally-derived phosphatidylcholine.
Neutral liposome compositions, for example, can be formed from
dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl
phosphatidylcholine (DPPC). Anionic liposome compositions generally
are formed from dimyristoyl phosphatidylglycerol, while anionic
fusogenic liposomes are formed primarily from dioleoyl
phosphatidylethanolamine (DOPE). Another type of liposomal
composition is formed from phosphatidylcholine (PC) such as, for
example, soybean PC, and egg PC. Another type is formed from
mixtures of phospholipid and/or phosphatidylcholine and/or
cholesterol.
[0172] Several studies have assessed the topical delivery of
liposomal drug formulations to the skin. Application of liposomes
containing interferon to guinea pig skin resulted in a reduction of
skin herpes sores while delivery of interferon via other means
(e.g. as a solution or as an emulsion) were ineffective (Weiner et
al., Journal of Drug Targeting, 1992, 2, 405-410). Further, an
additional study tested the efficacy of interferon administered as
part of a liposomal formulation to the administration of interferon
using an aqueous system, and concluded that the liposomal
formulation was superior to aqueous administration (du Plessis et
al., Antiviral Research, 1992, 18, 259-265).
[0173] Non-ionic liposomal systems have also been examined to
determine their utility in the delivery of drugs to the skin, in
particular systems comprising non-ionic surfactant and cholesterol.
Non-ionic liposomal formulations comprising Novasome.TM. I
(glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether)
and Novasome.TM. II (glyceryl
distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used
to deliver cyclosporin-A into the dermis of mouse skin. Results
indicated that such non-ionic liposomal systems were effective in
facilitating the deposition of cyclosporin-A into different layers
of the skin (Hu et al. S.T.P.Pharma. Sci., 1994, 4, 6, 466).
[0174] Liposomes also include "sterically stabilized" liposomes, a
term which, as used herein, refers to liposomes comprising one or
more specialized lipids that, when incorporated into liposomes,
result in enhanced circulation lifetimes relative to liposomes
lacking such specialized lipids. Examples of sterically stabilized
liposomes are those in which part of the vesicle-forming lipid
portion of the liposome (A) comprises one or more glycolipids, such
as monosialoganglioside G.sub.m1, or (B) is derivatized with one or
more hydrophilic polymers, such as a polyethylene glycol (PEG)
moiety. While not wishing to be bound by any particular theory, it
is thought in the art that, at least for sterically stabilized
liposomes containing gangliosides, sphingomyelin, or
PEG-derivatized lipids, the enhanced circulation half-life of these
sterically stabilized liposomes derives from a reduced uptake into
cells of the reticuloendothelial system (RES) (Allen et al., FEBS
Letters, 1987, 223, 42; Wu et al., Cancer Research, 1993, 53,
3765).
[0175] Various liposomes comprising one or more glycolipids are
known in the art. Papahadjopoulos et al. (Ann. N.Y. Acad. Sci.,
1987, 507, 64) reported the ability of monosialoganglioside
G.sub.m1, galactocerebroside sulfate and phosphatidylinositol to
improve blood half-lives of liposomes. These findings were
expounded upon by Gabizon et al. (Proc. Natl. Acad. Sci. U.S.A.,
1988, 85, 6949). U.S. Pat. No. 4,837,028 and WO 88/04924, both to
Allen et al., disclose liposomes comprising (1) sphingomyelin and
(2) the ganglioside G.sub.m1 or a galactocerebroside sulfate ester.
U.S. Pat. No. 5,543,152 (Webb et al.) discloses liposomes
comprising sphingomyelin. Liposomes comprising
1,2-sn-dimyristoylphosphat-idylcholine are disclosed in WO 97/13499
(Lim et al).
[0176] Many liposomes comprising lipids derivatized with one or
more hydrophilic polymers, and methods of preparation thereof, are
known in the art. Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53,
2778) described liposomes comprising a nonionic detergent,
2C.sub.1215G, that contains a PEG moiety. Illum et al. (FEBS Lett.,
1984, 167, 79) noted that hydrophilic coating of polystyrene
particles with polymeric glycols results in significantly enhanced
blood half-lives. Synthetic phospholipids modified by the
attachment of carboxylic groups of polyalkylene glycols (e.g., PEG)
are described by Sears (U.S. Pat. Nos. 4,426,330 and 4,534,899).
Klibanov et al. (FEBS Lett., 1990, 268, 235) described experiments
demonstrating that liposomes comprising phosphatidylethanolamine
(PE) derivatized with PEG or PEG stearate have significant
increases in blood circulation half-lives. Blume et al. (Biochimica
et Biophysica Acta, 1990, 1029, 91) extended such observations to
other PEG-derivatized phospholipids, e.g., DSPE-PEG, formed from
the combination of distearoylphosphatidylethanolamine (DSPE) and
PEG. Liposomes having covalently bound PEG moieties on their
external surface are described in European Patent No. EP 0 445 131
B1 and WO 90/04384 to Fisher. Liposome compositions containing 1-20
mole percent of PE derivatized with PEG, and methods of use
thereof, are described by Woodle et al. (U.S. Pat. Nos. 5,013,556
and 5,356,633) and Martin et al. (U.S. Pat. No. 5,213,804 and
European Patent No. EP 0 496 813 B1). Liposomes comprising a number
of other lipid-polymer conjugates are disclosed in WO 91/05545 and
U.S. Pat. No. 5,225,212 (both to Martin et al.) and in WO 94/20073
(Zalipsky et al.) Liposomes comprising PEG-modified ceramide lipids
are described in WO 96/10391 (Choi et al). U.S. Pat. No. 5,540,935
(Miyazaki et al.) and U.S. Pat. No. 5,556,948 (Tagawa et al.)
describe PEG-containing liposomes that can be further derivatized
with functional moieties on their surfaces.
[0177] A limited number of liposomes comprising nucleic acids are
known in the art. WO 96/40062 to Thierry et al. discloses methods
for encapsulating high molecular weight nucleic acids in liposomes.
U.S. Pat. No. 5,264,221 to Tagawa et al. discloses protein-bonded
liposomes and asserts that the contents of such liposomes may
include dsRNA. U.S. Pat. No. 5,665,710 to Rahman et al. describes
certain methods of encapsulating oligodeoxynucleotides in
liposomes. WO 97/04787 to Love et al. discloses liposomes
comprising dsRNAs targeted to the raf gene.
[0178] Transfersomes are yet another type of liposomes, and are
highly deformable lipid aggregates which are attractive candidates
for drug delivery vehicles. Transfersomes may be described as lipid
droplets which are so highly deformable that they are easily able
to penetrate through pores which are smaller than the droplet.
Transfersomes are adaptable to the environment in which they are
used, e.g. they are self-optimizing (adaptive to the shape of pores
in the skin), self-repairing, frequently reach their targets
without fragmenting, and often self-loading. To make transfersomes
it is possible to add surface edge-activators, usually surfactants,
to a standard liposomal composition. Transfersomes have been used
to deliver serum albumin to the skin. The transfersome-mediated
delivery of serum albumin has been shown to be as effective as
subcutaneous injection of a solution containing serum albumin.
[0179] Surfactants find wide application in formulations such as
emulsions (including microemulsions) and liposomes. The most common
way of classifying and ranking the properties of the many different
types of surfactants, both natural and synthetic, is by the use of
the hydrophile/lipophile balance (HLB). The nature of the
hydrophilic group (also known as the "head") provides the most
useful means for categorizing the different surfactants used in
formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel
Dekker, Inc., New York, N.Y., 1988, p. 285).
[0180] If the surfactant molecule is not ionized, it is classified
as a nonionic surfactant. Nonionic surfactants find wide
application in pharmaceutical and cosmetic products and are usable
over a wide range of pH values. In general their HLB values range
from 2 to about 18 depending on their structure. Nonionic
surfactants include nonionic esters such as ethylene glycol esters,
propylene glycol esters, glyceryl esters, polyglyceryl esters,
sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic
alkanolamides and ethers such as fatty alcohol ethoxylates,
propoxylated alcohols, and ethoxylated/propoxylated block polymers
are also included in this class. The polyoxyethylene surfactants
are the most popular members of the nonionic surfactant class.
[0181] If the surfactant molecule carries a negative charge when it
is dissolved or dispersed in water, the surfactant is classified as
anionic. Anionic surfactants include carboxylates such as soaps,
acyl lactylates, acyl amides of amino acids, esters of sulfuric
acid such as alkyl sulfates and ethoxylated alkyl sulfates,
sulfonates such as alkyl benzene sulfonates, acyl isethionates,
acyl taurates and sulfosuccinates, and phosphates. The most
important members of the anionic surfactant class are the alkyl
sulfates and the soaps.
[0182] If the surfactant molecule carries a positive charge when it
is dissolved or dispersed in water, the surfactant is classified as
cationic. Cationic surfactants include quaternary ammonium salts
and ethoxylated amines. The quaternary ammonium salts are the most
used members of this class.
[0183] If the surfactant molecule has the ability to carry either a
positive or negative charge, the surfactant is classified as
amphoteric. Amphoteric surfactants include acrylic acid
derivatives, substituted alkylamides, N-alkylbetaines and
phosphatides.
[0184] The use of surfactants in drug products, formulations and in
emulsions has been reviewed (Rieger, in Pharmaceutical Dosage
Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285).
[0185] Penetration Enhancers
[0186] In one embodiment, the present invention employs various
penetration enhancers to effect the efficient delivery of nucleic
acids, particularly dsRNAs, to the skin of animals. Most drugs are
present in solution in both ionized and nonionized forms. However,
usually only lipid soluble or lipophilic drugs readily cross cell
membranes. It has been discovered that even non-lipophilic drugs
may cross cell membranes if the membrane to be crossed is treated
with a penetration enhancer. In addition to aiding the diffusion of
non-lipophilic drugs across cell membranes, penetration enhancers
also enhance the permeability of lipophilic drugs.
[0187] Penetration enhancers may be classified as belonging to one
of five broad categories, i.e., surfactants, fatty acids, bile
salts, chelating agents, and non-chelating non-surfactants (Lee et
al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p.
92). Each of the above mentioned classes of penetration enhancers
are described below in greater detail.
[0188] Surfactants: In connection with the present invention,
surfactants (or "surface-active agents") are chemical entities
which, when dissolved in an aqueous solution, reduce the surface
tension of the solution or the interfacial tension between the
aqueous solution and another liquid, with the result that
absorption of dsRNAs through the mucosa is enhanced. In addition to
bile salts and fatty acids, these penetration enhancers include,
for example, sodium lauryl sulfate, polyoxyethylene-9-lauryl ether
and polyoxyethylene-20-cetyl ether) (Lee et al., Critical Reviews
in Therapeutic Drug Carrier Systems, 1991, p. 92); and
perfluorochemical emulsions, such as FC-43 (Takahashi et al., J.
Pharm. Pharmacol., 1988, 40, 252).
[0189] Fatty acids: Various fatty acids and their derivatives which
act as penetration enhancers include, for example, oleic acid,
lauric acid, capric acid (n-decanoic acid), myristic acid, palmitic
acid, stearic acid, linoleic acid, linolenic acid, dicaprate,
tricaprate, monoolein (1-monooleoyl-rac-glycerol), dilaurin,
caprylic acid, arachidonic acid, glycerol 1-monocaprate,
1-dodecylazacycloheptan-2-one, acylcarnitines, acylcholines,
C.sub.1-C.sub.10 alkyl esters thereof (e.g., methyl, isopropyl and
t-butyl), and mono- and di-glycerides thereof (i.e., oleate,
laurate, caprate, myristate, palmitate, stearate, linoleate, etc.)
(Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems,
1991, p. 92; Muranishi, Critical Reviews in Therapeutic Drug
Carrier Systems, 1990, 7, 1-33; El Hariri et al., J. Pharm.
Pharmacol., 1992, 44, 651-654).
[0190] Bile salts: The physiological role of bile includes the
facilitation of dispersion and absorption of lipids and fat-soluble
vitamins (Brunton, Chapter 38 in: Goodman & Gilman's The
Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al.
Eds., McGraw-Hill, New York, 1996, pp. 934-935). Various natural
bile salts, and their synthetic derivatives, act as penetration
enhancers. Thus the term "bile salts" includes any of the naturally
occurring components of bile as well as any of their synthetic
derivatives. The bile salts of the invention include, for example,
cholic acid (or its pharmaceutically acceptable sodium salt, sodium
cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic
acid (sodium deoxycholate), glucholic acid (sodium glucholate),
glycholic acid (sodium glycocholate), glycodeoxycholic acid (sodium
glycodeoxycholate), taurocholic acid (sodium taurocholate),
taurodeoxycholic acid (sodium taurodeoxycholate), chenodeoxycholic
acid (sodium chenodeoxycholate), ursodeoxycholic acid (UDCA),
sodium tauro-24,25-dihydro-fusidate (STDHF), sodium
glycodihydrofusidate and polyoxyethylene-9-lauryl ether (POE) (Lee
et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991,
page 92; Swinyard, Chapter 39 In: Remington's Pharmaceutical
Sciences, 18th Ed., Gennaro, ed., Mack Publishing Co., Easton, Pa.,
1990, pages 782-783; Muranishi, Critical Reviews in Therapeutic
Drug Carrier Systems, 1990, 7, 1-33; Yamamoto et al., J. Pharm.
Exp. Ther., 1992, 263, 25; Yamashita et al., J. Pharm. Sci., 1990,
79, 579-583).
[0191] Chelating Agents: Chelating agents, as used in connection
with the present invention, can be defined as compounds that remove
metallic ions from solution by forming complexes therewith, with
the result that absorption of dsRNAs through the mucosa is
enhanced. With regards to their use as penetration enhancers in the
present invention, chelating agents have the added advantage of
also serving as DNase inhibitors, as most characterized DNA
nucleases require a divalent metal ion for catalysis and are thus
inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618,
315-339). Chelating agents of the invention include but are not
limited to disodium ethylenediaminetetraacetate (EDTA), citric
acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and
homovanilate), N-acyl derivatives of collagen, laureth-9 and
N-amino acyl derivatives of beta-diketones (enamines)(Lee et al.,
Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page
92; Muranishi, Critical Reviews in Therapeutic Drug Carrier
Systems, 1990, 7, 1-33; Buur et al., J. Control Rel., 1990, 14,
43-51).
[0192] Non-chelating non-surfactants: As used herein, non-chelating
non-surfactant penetration enhancing compounds can be defined as
compounds that demonstrate insignificant activity as chelating
agents or as surfactants but that nonetheless enhance absorption of
dsRNAs through the alimentary mucosa (Muranishi, Critical Reviews
in Therapeutic Drug Carrier Systems, 1990, 7, 1-33). This class of
penetration enhancers include, for example, unsaturated cyclic
ureas, 1-alkyl- and 1-alkenylazacyclo-alkanone derivatives (Lee et
al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991,
page 92); and non-steroidal anti-inflammatory agents such as
diclofenac sodium, indomethacin and phenylbutazone (Yamashita et
al., J. Pharm. Pharmacol., 1987, 39, 621-626).
[0193] Agents that enhance uptake of dsRNAs at the cellular level
may also be added to the pharmaceutical and other compositions of
the present invention. For example, cationic lipids, such as
lipofectin (Junichi et al, U.S. Pat. No. 5,705,188), cationic
glycerol derivatives, and polycationic molecules, such as
polylysine (Lollo et al., PCT Application WO 97/30731), are also
known to enhance the cellular uptake of dsRNAs.
[0194] Other agents may be utilized to enhance the penetration of
the administered nucleic acids, including glycols such as ethylene
glycol and propylene glycol, pyrrols such as 2-pyrrol, azones, and
terpenes such as limonene and menthone.
[0195] Carriers
[0196] Certain compositions of the present invention also
incorporate carrier compounds in the formulation. As used herein,
"carrier compound" or "carrier" can refer to a nucleic acid, or
analog thereof, which is inert (i.e., does not possess biological
activity per se) but is recognized as a nucleic acid by in vivo
processes that reduce the bioavailability of a nucleic acid having
biological activity by, for example, degrading the biologically
active nucleic acid or promoting its removal from circulation. The
coadministration of a nucleic acid and a carrier compound,
typically with an excess of the latter substance, can result in a
substantial reduction of the amount of nucleic acid recovered in
the liver, kidney or other extracirculatory reservoirs, presumably
due to competition between the carrier compound and the nucleic
acid for a common receptor. For example, the recovery of a
partially phosphorothioate dsRNA in hepatic tissue can be reduced
when it is coadministered with polyinosinic acid, dextran sulfate,
polycytidic acid or
4-acetamido-4'isothiocyano-stilbene-2,2'-disulfonic acid (Miyao et
al., Antisense Res. Dev., 1995, 5, 115-121; Takakura et al.,
Antisense & Nucl. Acid Drug Dev., 1996, 6, 177-183.
[0197] Excipients
[0198] In contrast to a carrier compound, a "pharmaceutical
carrier" or "excipient" is a pharmaceutically acceptable solvent,
suspending agent or any other pharmacologically inert vehicle for
delivering one or more nucleic acids to an animal. The excipient
may be liquid or solid and is selected, with the planned manner of
administration in mind, so as to provide for the desired bulk,
consistency, etc., when combined with a nucleic acid and the other
components of a given pharmaceutical composition. Typical
pharmaceutical carriers include, but are not limited to, binding
agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or
hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and
other sugars, microcrystalline cellulose, pectin, gelatin, calcium
sulfate, ethyl cellulose, polyacrylates or calcium hydrogen
phosphate, etc.); lubricants (e.g., magnesium stearate, talc,
silica, colloidal silicon dioxide, stearic acid, metallic
stearates, hydrogenated vegetable oils, corn starch, polyethylene
glycols, sodium benzoate, sodium acetate, etc.); disintegrants
(e.g., starch, sodium starch glycolate, etc.); and wetting agents
(e.g., sodium lauryl sulphate, etc).
[0199] Pharmaceutically acceptable organic or inorganic excipient
suitable for non-parenteral administration which do not
deleteriously react with nucleic acids can also be used to
formulate the compositions of the present invention. Suitable
pharmaceutically acceptable carriers include, but are not limited
to, water, salt solutions, alcohols, polyethylene glycols, gelatin,
lactose, amylose, magnesium stearate, talc, silicic acid, viscous
paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and the
like.
[0200] Formulations for topical administration of nucleic acids may
include sterile and non-sterile aqueous solutions, non-aqueous
solutions in common solvents such as alcohols, or solutions of the
nucleic acids in liquid or solid oil bases. The solutions may also
contain buffers, diluents and other suitable additives.
Pharmaceutically acceptable organic or inorganic excipients
suitable for non-parenteral administration which do not
deleteriously react with nucleic acids can be used.
[0201] Suitable pharmaceutically acceptable excipients include, but
are not limited to, water, salt solutions, alcohol, polyethylene
glycols, gelatin, lactose, amylose, magnesium stearate, talc,
silicic acid, viscous paraffin, hydroxymethylcellulose,
polyvinylpyrrolidone and the like.
[0202] Pharmaceutical Compositions for the Delivery to the
Respiratory Tract
[0203] Another aspect of the invention provides for the delivery of
IRNA agents to the respiratory tract, particularly for the
treatment of cystic fibrosis. The respiratory tract includes the
upper airways, including the oropharynx and larynx, followed by the
lower airways, which include the trachea followed by bifurcations
into the bronchi and bronchioli. The upper and lower airways are
called the conductive airways. The terminal bronchioli then divide
into respiratory bronchioli which then lead to the ultimate
respiratory zone, the alveoli, or deep lung. The deep lung, or
alveoli, are the primary target of inhaled therapeutic aerosols for
systemic delivery of iRNA agents.
[0204] Pulmonary delivery compositions can be delivered by
inhalation by the patient of a dispersion so that the composition,
preferably the iRNA agent, within the dispersion can reach the lung
where it can, for example, be readily absorbed through the alveolar
region directly into blood circulation. Pulmonary delivery can be
effective both for systemic delivery and for localized delivery to
treat diseases of the lungs.
[0205] Pulmonary delivery can be achieved by different approaches,
including the use of nebulized, aerosolized, micellular and dry
powder-based formulations; administration by inhalation may be oral
and/or nasal. Delivery can be achieved with liquid nebulizers,
aerosol-based inhalers, and dry powder dispersion devices.
Metered-dose devices are preferred. One of the benefits of using an
atomizer or inhaler is that the potential for contamination is
minimized because the devices are self contained. Dry powder
dispersion devices, for example, deliver drugs that may be readily
formulated as dry powders. An iRNA composition may be stably stored
as lyophilized or spray-dried powders by itself or in combination
with suitable powder carriers. The delivery of a composition for
inhalation can be mediated by a dosing timing element which can
include a timer, a dose counter, time measuring device, or a time
indicator which when incorporated into the device enables dose
tracking, compliance monitoring, and/or dose triggering to a
patient during administration of the aerosol medicament.
[0206] Examples of pharmaceutical devices for aerosol delivery
include metered dose inhalers (MDIs), dry powder inhalers (DPIs),
and air-jet nebulizers. Exemplary delivery systems by inhalation
which can be readily adapted for delivery of the subject iRNA
agents are described in, for example, U.S. Pat. Nos. 5,756,353;
5,858,784; and PCT applications WO98/31346; WO98/10796; WO00/27359;
WO01/54664; WO02/060412. Other aerosol formulations that may be
used for delivering the iRNA agents are described in U.S. Pat. Nos.
6,294,153; 6,344,194; 6,071,497, and PCT applications WO02/066078;
WO02/053190; WO01/60420; WO00/66206. Further, methods for
delivering iRNA agents can be adapted from those used in delivering
other oligonucleotides (e.g., an antisense oligonucleotide) by
inhalation, such as described in Templin et al., Antisense Nucleic
Acid Drug Dev, 2000, 10:359-68; Sandrasagra et al., Expert Opin
Biol Ther, 2001, 1:979-83; Sandrasagra et al., Antisense Nucleic
Acid Drug Dev, 2002, 12:177-81.
[0207] The delivery of the inventive agents may also involve the
administration of so called "pro-drugs", i.e. formulations or
chemical modifications of a therapeutic substance that require some
form of processing or transport by systems innate to the subject
organism to release the therapeutic substance, preferably at the
site where its action is desired; this latter embodiment may be
used in conjunction with delivery of the respiratory tract, but
also together with other embodiments of the present invention. For
example, the human lungs can remove or rapidly degrade
hydrolytically cleavable deposited aerosols over periods ranging
from minutes to hours. In the upper airways, ciliated epithelia
contribute to the "mucociliary excalator" by which particles are
swept from the airways toward the mouth. Pavia, D., "Lung
Mucociliary Clearance," in Aerosols and the Lung: Clinical and
Experimental Aspects, Clarke, S. W. and Pavia, D., Eds.,
Butterworths, London, 1984. In the deep lungs, alveolar macrophages
are capable of phagocytosing particles soon after their deposition.
Warheit et al. Microscopy Res. Tech., 26: 412-422 (1993); and
Brain, J. D., "Physiology and Pathophysiology of Pulmonary
Macrophages," in The Reticuloendothelial System, S. M. Reichard and
J. Filkins, Eds., Plenum, New. York., pp. 315-327, 1985.
[0208] In preferred embodiments, particularly where systemic dosing
with the iRNA agent is desired, the aerosoled iRNA agents are
formulated as microparticles. Microparticles having a diameter of
between 0.5 and ten microns can penetrate the lungs, passing
through most of the natural barriers. A diameter of less than ten
microns is required to bypass the throat; a diameter of 0.5 microns
or greater is required to avoid being exhaled.
[0209] Other Components
[0210] The compositions of the present invention may additionally
contain other adjunct components conventionally found in
pharmaceutical compositions, at their art-established usage levels.
Thus, for example, the compositions may contain additional,
compatible, pharmaceutically-active materials such as, for example,
antipruritics, astringents, local anesthetics or anti-inflammatory
agents, or may contain additional materials useful in physically
formulating various dosage forms of the compositions of the present
invention, such as dyes, flavoring agents, preservatives,
antioxidants, opacifiers, thickening agents and stabilizers.
However, such materials, when added, should not unduly interfere
with the biological activities of the components of the
compositions of the present invention. The formulations can be
sterilized and, if desired, mixed with auxiliary agents, e.g.,
lubricants, preservatives, stabilizers, wetting agents,
emulsifiers, salts for influencing osmotic pressure, buffers,
colorings, flavorings and/or aromatic substances and the like which
do not deleteriously interact with the nucleic acid(s) of the
formulation.
[0211] Aqueous suspensions may contain substances which increase
the viscosity of the suspension including, for example, sodium
carboxymethylcellulose, sorbitol and/or dextran. The suspension may
also contain stabilizers.
[0212] Certain embodiments of the invention provide pharmaceutical
compositions containing (a) one or more dsRNA agents and (b) one or
more other chemotherapeutic agents which function by a non-RNA
interference mechanism. Examples of such chemotherapeutic agents
include but are not limited to daunorubicin, daunomycin,
dactinomycin, doxorubicin, epirubicin, idarubicin, esorubicin,
bleomycin, mafosfamide, ifosfamide, cytosine arabinoside,
bis-chloroethylnitrosurea, busulfan, mitomycin C, actinomycin D,
mithramycin, prednisone, hydroxyprogesterone, testosterone,
tamoxifen, dacarbazine, procarbazine, hexamethylmelamine,
pentamethylmelamine, mitoxantrone, amsacrine, chlorambucil,
methylcyclohexylnitrosurea, nitrogen mustards, melphalan,
cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine,
5-azacytidine, hydroxyurea, deoxycoformycin,
4-hydroxyperoxycyclophosphoramide, 5-fluorouracil (5-FU),
5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine,
taxol, vincristine, vinblastine, etoposide (VP-16), trimetrexate,
irinotecan, topotecan, gemcitabine, teniposide, cisplatin and
diethylstilbestrol (DES). See, generally, The Merck Manual of
Diagnosis and Therapy, 15th Ed. 1987, pp. 1206-1228, Berkow et al.,
eds., Rahway, N.J. When used with the compounds of the invention,
such chemotherapeutic agents may be used individually (e.g., 5-FU
and oligonucleotide), sequentially (e.g., 5-FU and oligonucleotide
for a period of time followed by MTX and oligonucleotide), or in
combination with one or more other such chemotherapeutic agents
(e.g., 5-FU, MTX and oligonucleotide, or 5-FU, radiotherapy and
oligonucleotide). Anti-inflammatory drugs, including but not
limited to nonsteroidal anti-inflammatory drugs and
corticosteroids, and antiviral drugs, including but not limited to
ribivirin, vidarabine, acyclovir and ganciclovir, may also be
combined in compositions of the invention. See, generally, The
Merck Manual of Diagnosis and Therapy, 15th Ed., Berkow et al.,
eds., 1987, Rahway, N.J., pages 2499-2506 and 46-49, respectively).
Other non-dsRNA chemotherapeutic agents are also within the scope
of this invention. Two or more combined compounds may be used
together or sequentially.
[0213] Toxicity and therapeutic efficacy of such compounds can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., for determining the LD50 (the dose
lethal to 50% of the population) and the ED50 (the dose
therapeutically effective in 50% of the population). The dose ratio
between toxic and therapeutic effects is the therapeutic index and
it can be expressed as the ratio LD50/ED50. Compounds which exhibit
high therapeutic indices are preferred.
[0214] The data obtained from cell culture assays and animal
studies can be used in formulation a range of dosage for use in
humans. The dosage of compositions of the invention lies generally
within a range of circulating concentrations that include the ED50
with little or no toxicity. The dosage may vary within this range
depending upon the dosage form employed and the route of
administration utilized. For any compound used in the method of the
invention, the therapeutically effective dose can be estimated
initially from cell culture assays. A dose may be formulated in
animal models to achieve a circulating plasma concentration range
of the compound or, when appropriate, of the polypeptide product of
a target sequence (e.g., achieving a decreased concentration of the
polypeptide) that includes the IC50 (i.e., the concentration of the
test compound which achieves a half-maximal inhibition of symptoms)
as determined in cell culture. Such information can be used to more
accurately determine useful doses in humans. Levels in plasma may
be measured, for example, by high performance liquid
chromatography.
[0215] In addition to their administration individually or as a
plurality, as discussed above, the dsRNAs of the invention can be
administered in combination with other known agents effective in
treatment of influenza infection. In any event, the administering
physician can adjust the amount and timing of dsRNA administration
on the basis of results observed using standard measures of
efficacy known in the art or described herein.
[0216] Treatment Methods and Routes of Delivery
[0217] A composition that includes an iRNA agent, e.g., an iRNA
agent that targets influenza virus, can be delivered to a subject
by a variety of routes to achieve either local delivery to the site
of action of systemic delivery to the subject. Exemplary routes
include direct local administration to the site of treatment, such
as the lungs and nasal passage as well as intravenous, nasal, oral,
and ocular delivery. The preferred means of administering the iRNA
agents of the present invention is through direct administration to
the lungs and nasal passage as a liquid, aerosol or nubulized
solution.
[0218] In general, the delivery of the iRNA agents of the present
invention is done to achieve delivery into the subject to the site
of infection. The preferred means of achieving this is through
either a local administration to the lungs or nasal passage, e.g.
into the respiratory tissues via inhalation or intranasal
administration, or via systemic administration, e.g. parental
administration.
[0219] Formulations for inhalation or parenteral administration are
well known in the art. Such formulation may include sterile aqueous
solutions which may also contain buffers, diluents and other
suitable additives. For intravenous use, the total concentration of
solutes should be controlled to render the preparation
isotonic.
[0220] The active compounds disclosed herein are preferably
administered to the lung(s) or nasal passage of a subject by any
suitable means. Active compounds may be administered by
administering an aerosol suspension of respirable particles
comprised of the active compound or active compounds, which the
subject inhales. The active compound can be aerosolized in a
variety of forms, such as, but not limited to, dry powder
inhalants, metered dose inhalants, or liquid/liquid suspensions.
The respirable particles may be liquid or solid. The particles may
optionally contain other therapeutic ingredients such as amiloride,
benzamil or phenamil, with the selected compound included in an
amount effective to inhibit the reabsorption of water from airway
mucous secretions, as described in U.S. Pat. No. 4,501,729.
[0221] The particulate pharmaceutical composition may optionally be
combined with a carrier to aid in dispersion or transport. A
suitable carrier such as a sugar (i.e., lactose, sucrose,
trehalose, mannitol) may be blended with the active compound or
compounds in any suitable ratio (e.g., a 1 to 1 ratio by
weight).
[0222] Particles comprised of the active compound for practicing
the present invention should include particles of respirable size,
that is, particles of a size sufficiently small to pass through the
mouth or nose and larynx upon inhalation and into the bronchi and
alveoli of the lungs. In general, particles ranging from about 1 to
10 microns in size (more particularly, less than about 5 microns in
size) are respirable. Particles of non-respirable size which are
included in the aerosol tend to deposit in the throat and be
swallowed, and the quantity of non-respirable particles in the
aerosol is preferably minimized. For nasal administration, a
particle size in the range of 10-500 uM is preferred to ensure
retention in the nasal cavity.
[0223] Liquid pharmaceutical compositions of active compound for
producing an aerosol may be prepared by combining the active
compound with a suitable vehicle, such as sterile pyrogen free
water. The hypertonic saline solutions used to carry out the
present invention are preferably sterile, pyrogen-free solutions,
comprising from one to fifteen percent (by weight) of the
physiologically acceptable salt, and more preferably from three to
seven percent by weight of the physiologically acceptable salt.
[0224] Aerosols of liquid particles comprising the active compound
may be produced by any suitable means, such as with a
pressure-driven jet nebulizer or an ultrasonic nebulizer. See,
e.g., U.S. Pat. No. 4,501,729. Nebulizers are commercially
available devices which transform solutions or suspensions of the
active ingredient into a therapeutic aerosol mist either by means
of acceleration of compressed gas, typically air or oxygen, through
a narrow venturi orifice or by means of ultrasonic agitation.
[0225] Suitable formulations for use in nebulizers consist of the
active ingredient in a liquid carrier, the active ingredient
comprising up to 40% w/w of the formulation, but preferably less
than 20% w/w. The carrier is typically water (and most preferably
sterile, pyrogen-free water) or a dilute aqueous alcoholic
solution, preferably made isotonic, but may be hypertonic with body
fluids by the addition of, for example, sodium chloride. Optional
additives include preservatives if the formulation is not made
sterile, for example, methyl hydroxybenzoate, antioxidants,
flavoring agents, volatile oils, buffering agents and
surfactants.
[0226] Aerosols of solid particles comprising the active compound
may likewise be produced with any solid particulate therapeutic
aerosol generator. Aerosol generators for administering solid
particulate therapeutics to a subject produce particles which are
respirable and generate a volume of aerosol containing a
predetermined metered dose of a therapeutic at a rate suitable for
human administration. One illustrative type of solid particulate
aerosol generator is an insufflator. Suitable formulations for
administration by insufflation include finely comminuted powders
which may be delivered by means of an insufflator or taken into the
nasal cavity in the manner of a snuff. In the insufflator, the
powder (e.g., a metered dose thereof effective to carry out the
treatments described herein) is contained in capsules or
cartridges, typically made of gelatin or plastic, which are either
pierced or opened in situ and the powder delivered by air drawn
through the device upon inhalation or by means of a
manually-operated pump. The powder employed in the insufflator
consists either solely of the active ingredient or of a powder
blend comprising the active ingredient, a suitable powder diluent,
such as lactose, and an optional surfactant. The active ingredient
typically comprises from 0.1 to 100 w/w of the formulation.
[0227] A second type of illustrative aerosol generator comprises a
metered dose inhaler. Metered dose inhalers are pressurized aerosol
dispensers, typically containing a suspension or solution
formulation of the active ingredient in a liquefied propellant.
During use these devices discharge the formulation through a valve
adapted to deliver a metered volume, typically from 10 to 200 ul,
to produce a fine particle spray containing the active ingredient.
Suitable propellants include certain chlorofluorocarbon compounds,
for example, dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane and mixtures thereof. The formulation may
additionally contain one or more co-solvents, for example, ethanol,
surfactants, such as oleic acid or sorbitan trioleate, antioxidant
and suitable flavoring agents.
[0228] An iRNA agent can be incorporated into pharmaceutical
compositions suitable for administration. For example, compositions
can include one or more species of an iRNA agent and a
pharmaceutically acceptable carrier. As used herein the language
"pharmaceutically acceptable carrier" is intended to include any
and all solvents, dispersion media, coatings, antibacterial and
antifungal agents, isotonic and absorption delaying agents, and the
like, compatible with pharmaceutical administration. The use of
such media and agents for pharmaceutically active substances is
well known in the art. Except insofar as any conventional media or
agent is incompatible with the active compound, use thereof in the
compositions is contemplated. Supplementary active compounds can
also be incorporated into the compositions.
[0229] Administration can be provided by the subject or by another
person, e.g., a caregiver. A caregiver can be any entity involved
with providing care to the human: for example, a hospital, hospice,
doctor's office, outpatient clinic; a healthcare worker such as a
doctor, nurse, or other practitioner; or a spouse or guardian, such
as a parent. The medication can be provided in measured doses or in
a dispenser which delivers a metered dose.
[0230] The term "therapeutically effective amount" is the amount
present in the composition that is needed to provide the desired
level of drug in the subject to be treated to give the anticipated
physiological response.
[0231] The term "physiologically effective amount" is that amount
delivered to a subject to give the desired palliative or curative
effect.
[0232] The term "pharmaceutically acceptable carrier" means that
the carrier can be taken into the lungs with no significant adverse
toxicological effects on the lungs.
[0233] The term "co-administration" refers to administering to a
subject two or more agents, and in particular two or more iRNA
agents. The agents can be contained in a single pharmaceutical
composition and be administered at the same time, or the agents can
be contained in separate formulation and administered serially to a
subject. So long as the two agents can be detected in the subject
at the same time, the two agents are said to be
co-administered.
[0234] The types of pharmaceutical excipients that are useful as
carrier include stabilizers such as human serum albumin (HSA),
bulking agents such as carbohydrates, amino acids and polypeptides;
pH adjusters or buffers; salts such as sodium chloride; and the
like. These carriers may be in a crystalline or amorphous form or
may be a mixture of the two.
[0235] Bulking agents that are particularly valuable include
compatible carbohydrates, polypeptides, amino acids or combinations
thereof. Suitable carbohydrates include monosaccharides such as
galactose, D-mannose, sorbose, and the like; disaccharides, such as
lactose, trehalose, and the like; cyclodextrins, such as
2-hydroxypropyl-.beta.-cyclodextrin; and polysaccharides, such as
raffinose, maltodextrins, dextrans, and the like; alditols, such as
mannitol, xylitol, and the like. A preferred group of carbohydrates
includes lactose, threhalose, raffinose maltodextrins, and
mannitol. Suitable polypeptides include aspartame. Amino acids
include alanine and glycine, with glycine being preferred.
[0236] Suitable pH adjusters or buffers include organic salts
prepared from organic acids and bases, such as sodium citrate,
sodium ascorbate, and the like; sodium citrate is preferred.
[0237] Dosage
[0238] An iRNA agent can be administered at a unit dose less than
about 75 mg per kg of bodyweight, or less than about 70, 60, 50,
40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001, or
0.0005 mg per kg of bodyweight, and less than 200 nmol of iRNA
agent (e.g., about 4.4.times.1016 copies) per kg of bodyweight, or
less than 1500, 750, 300, 150, 75, 15, 7.5, 1.5, 0.75, 0.15, 0.075,
0.015, 0.0075, 0.0015, 0.00075, 0.00015 nmol of iRNA agent per kg
of bodyweight. The unit dose, for example, can be administered by
injection (e.g., intravenous or intramuscular, intrathecally, or
directly into an organ), an inhaled dose, or a topical
application.
[0239] Delivery of an iRNA agent directly to an organ (e.g., to the
lung) can be at a dosage on the order of about 0.00001 mg to about
3 mg per organ, or preferably about 0.0001-0.001 mg per organ,
about 0.03-3.0 mg per organ, about 0.1-3.0 mg per eye or about
0.3-3.0 mg per organ.
[0240] The dosage can be an amount effective to treat or prevent a
disease or disorder. It can be given prophylactically or as the
primary or a part of a treatment protocol.
[0241] In one embodiment, the unit dose is administered less
frequently than once a day, e.g., less than every 2, 4, 8 or 30
days. In another embodiment, the unit dose is not administered with
a frequency (e.g., not a regular frequency). For example, the unit
dose may be administered a single time. Because iRNA agent mediated
silencing can persist for several days after administering the iRNA
agent composition, in many instances, it is possible to administer
the composition with a frequency of less than once per day, or, for
some instances, only once for the entire therapeutic regimen.
[0242] In one embodiment, a subject is administered an initial
dose, and one or more maintenance doses of an iRNA agent, e.g., a
double-stranded iRNA agent, or siRNA agent, (e.g., a precursor,
e.g., a larger iRNA agent which can be processed into an siRNA
agent, or a DNA which encodes an iRNA agent, e.g., a
double-stranded iRNA agent, or siRNA agent, or precursor thereof).
The maintenance dose or doses are generally lower than the initial
dose, e.g., one-half less of the initial dose. A maintenance
regimen can include treating the subject with a dose or doses
ranging from 0.01 to 75 mg/kg of body weight per day, e.g., 70, 60,
50, 40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001, or
0.0005 mg per kg of body weight per day. The maintenance doses are
preferably administered no more than once every 5, 10, or 30 days.
Further, the treatment regimen may last for a period of time which
will vary depending upon the nature of the particular disease, its
severity and the overall condition of the patient. In preferred
embodiments the dosage may be delivered no more than once per day,
e.g., no more than once per 24, 36, 48, or more hours, e.g., no
more than once every 5 or 8 days. Following treatment, the patient
can be monitored for changes in his condition and for alleviation
of the symptoms of the disease state. The dosage of the compound
may either be increased in the event the patient does not respond
significantly to current dosage levels, or the dose may be
decreased if an alleviation of the symptoms of the disease state is
observed, if the disease state has been ablated, or if undesired
side-effects are observed.
[0243] The effective dose can be administered in a single dose or
in two or more doses, as desired or considered appropriate under
the specific circumstances. If desired to facilitate repeated or
frequent infusions, implantation of a delivery device, e.g., a
pump, semi-permanent stent (e.g., intravenous, intraperitoneal,
intracisternal or intracapsular), or reservoir may be
advisable.
[0244] Following successful treatment, it may be desirable to have
the patient undergo maintenance therapy to prevent the recurrence
of the disease state, wherein the compound of the invention is
administered in maintenance doses, ranging from 0.001 g to 100 g
per kg of body weight (see U.S. Pat. No. 6,107,094).
[0245] The concentration of the iRNA agent composition is an amount
sufficient to be effective in treating or preventing a disorder or
to regulate a physiological condition in humans. The concentration
or amount of iRNA agent administered will depend on the parameters
determined for the agent and the method of administration, e.g.
nasal, buccal, or pulmonary. For example, nasal formulations tend
to require much lower concentrations of some ingredients in order
to avoid irritation or burning of the nasal passages. It is
sometimes desirable to dilute an oral formulation up to 10-100
times in order to provide a suitable nasal formulation.
[0246] Certain factors may influence the dosage required to
effectively treat a subject, including but not limited to the
severity of the disease or disorder, previous treatments, the
general health and/or age of the subject, and other diseases
present. It will also be appreciated that the effective dosage of
an iRNA agent such as an siRNA used for treatment may increase or
decrease over the course of a particular treatment. Changes in
dosage may result and become apparent from the results of
diagnostic assays. For example, the subject can be monitored after
administering an iRNA agent composition. Based on information from
the monitoring, an additional amount of the iRNA agent composition
can be administered.
[0247] Dosing is dependent on severity and responsiveness of the
disease condition to be treated, with the course of treatment
lasting from several days to several months, or until a cure is
effected or a diminution of disease state is achieved. Optimal
dosing schedules can be calculated from measurements of drug
accumulation in the body of the patient. Persons of ordinary skill
can easily determine optimum dosages, dosing methodologies and
repetition rates. Optimum dosages may vary depending on the
relative potency of individual compounds, and can generally be
estimated based on EC50s found to be effective in in vitro and in
vivo animal models as described above.
[0248] The invention is further illustrated by the following
examples, which should not be construed as further limiting.
EXAMPLES
[0249] Nucleic acid sequences are represented below using standard
nomenclature, and specifically the abbreviations of Table 3.
TABLE-US-00011 TABLE 3 Abbreviations of nucleotide monomers used in
nucleic acid sequence representation. It will be understood that
these monomers, when present in an oligonucleotide, are mutually
linked by 5'-3'-phosphodiester bonds. Abbreviation.sup.a
Nucleotide(s) A, a 2'-deoxy-adenosine-5'-phosphate,
adenosine-5'-phosphate C, c 2'-deoxy-cytidine-5'-phosphate,
cytidine-5'-phosphate G, g 2'-deoxy-guanosine-5'-phosphate,
guanosine-5'-phosphate T, t 2'-deoxy-thymidine-5'-phosphate,
thymidine-5'-phosphate U, u 2'-deoxy-uridine-5'-phosphate,
uridine-5'-phosphate N, n any 2'-deoxy-nucleotide/nucleotide (G, A,
C, or T, g, a, c or u) am 2'-O-methyladenosine-5'-phosphate cm
2'-O-methylcytidine-5'-phosphate gm
2'-O-methylguanosine-5'-phosphate tm
2'-O-methyl-thymidine-5'-phosphate um
2'-O-methyluridine-5'-phosphate A, C, G, T, U, underlined:
nucleoside-5'-phosphorothioate a, c, g, t, u x universal base
.sup.acapital letters represent 2'-deoxyribonucleotides (DNA),
lower case letters represent ribonucleotides (RNA)
[0250] Source of Reagents
[0251] Where the source of a reagent is not specifically given
herein, such reagent may be obtained from any supplier of reagents
for molecular biology at a quality/purity standard for application
in molecular biology.
Example 1
Selection of Sequences
[0252] siRNA design was carried out to identify siRNAs targeting
Influenza A mRNAs of MP, NP, PA, PB1 and PB2 protein. In a first
round, the siRNA in silico selection resulted in 44 sequences
targeting MP, 3 sequences targeting NP and 1 sequence targeting
PB1. No siRNAs specific for influenza A genes PA or PB2 passed the
first selection process demanding 80% target coverage and 80%
target efficiency (see below).
[0253] To setup an environment for sequence analysis, the fastA
package (Pearson, W. R., & Lipman, D. J., PNAS 1988, 85:2444)
was downloaded from ftp://ftp.virginia.edu/pub/ and installed on a
workstation under the Suse Linux.RTM. 9.3 operating system with
standard installation settings. For the purpose of running perl
scripts, it was ensured that the perl interpreter, version 5.8.6
(copyright 1987-2004, Larry Wall), coming with the Suse Linux 9.3
standard installation was functional. BioEdit Sequence Alignment
Editor (Hall, T. A., Nucl. Acids. Symp. Ser. 1999, 41:95) was
downloaded from the Brown Lab Web Server at the web site of North
Carolina State University and installed on a computer under
Microsoft Windows2000.RTM. operating system.
[0254] Workflow for the in silico selection was as follows:
influenza A sequences of interest were downloaded, aligned and a
statistics was generated to obtain distribution of bases at every
position relative to a calculated consensus. A perl script was used
to identify candidate target regions satisfying defined cut-off
criteria. siRNA sequences to candidate target regions were analyzed
for specificity by fasta algorithm to human RefSeq database.
Another perl script was used to score siRNAs according to predicted
specificity. Finally, those siRNAs were manually selected that
satisfied specificity criteria.
[0255] Influenza A sequences of interest available on Jun. 24, 2005
were downloaded from NCBI Influenza Virus Database available on the
web site of the National Center for Biotechnology Information. The
number of sequences per gene is shown for MP, NP, PB1, PB2, and PA
in Table 4, the corresponding accession numbers are given in Table
4. TABLE-US-00012 TABLE 4 Number of gene sequences for influenza
genes MP, NP PB1, PB2, and PA from various viral subtypes that were
employed in in silico selection of siRNA sequences Gene H1N1 H2N2
H3N2 H5N1 H7N3 H7N7 H9N2 Total MP 10 2 13 166 28 16 128 363 NP 12 3
10 169 12 6 138 350 PB1 3 2 10 163 10 7 127 322 PB2 2 1 10 164 12 9
133 331 PA 2 2 10 171 11 8 124 328
[0256] The ClustalW multiple alignment function (Thompson, J. D.,
et al., Nucleic Acids Res. 1994; 22:4673) of BioEdit Sequence
Alignment Editor was used to generate a global alignment of all
sequences using default parameters for each target, respectively. A
Positional nucleotide numerical summary output was generated
providing information on base distribution at every position
relative to the calculated consensus sequence for each target.
[0257] Cut-off criteria for the identification of candidate
targeting regions of 19 nucleotides in length were defined as:
[0258] Criterium 1, target coverage: at least 80% of all sequences
available for the respective influenza A gene needed to be
represented in a candidate region
[0259] Criterium 2, targeting efficiency: at least 80% of all
sequences in which the candidate region was represented needed to
be identical within the candidate region.
[0260] Criterium 1 was defined in order to avoid regions for which
little sequence information was available, criterium 2 ensures
targeting of a high number of subtypes.
[0261] A perl script was used for screening the Positional
nucleotide numerical summary file to identify candidate target
regions with a length of 19 bases matching the cut-off criteria and
to generate a file to be used as fastA input in the following
analysis step. For script input the total number of sequences were
entered for each target and a value of 80 for percentage
conservation. All candidate sense siRNA sequences corresponding to
the most frequent sequences in the candidate target regions were
extracted and saved in a fastA-formatted file. In order to consider
potential dTdT-overhang interactions of siRNAs with the target
sequence, all sequences were extended at the 5' end with `AA`
resulting in 21mer input sequences. A further file was generated
for each candidate target region with information on region
properties: target coverage (sequences present) targeting
efficiency, total number of mismatches, number of conserved
sequences, and number of sequences present.
[0262] For further selection, candidate siRNAs were ranked
according to their predicted potential for interacting with host
(here, without limitation, human) genes (off-target potential).
siRNAs with low off-target potential are assumed to be more
specific in vivo.
[0263] For predicting siRNA-specific off-target potential, the
following assumptions were made:
[0264] 1) positions 2 to 9 (counting 5' to 3') of a strand (seed
region) contributes more to the off-target potential than the
remaining sequence (non-seed region) (Haley, B., and Zamore, P. D.,
Nat Struct Mol. Biol. 2004, 11:599).
[0265] 2) an off-target score can be calculated for each hit, based
on identity to siRNA sequence and position of mismatches
[0266] 3) by introducing appropriate nucleotide modifications into
the sense strand (e.g. all nucleotides comprising a pyrimidine base
are 2'-O-methyl modified nucleotides), the sense strand can be made
inactive towards RNA interference; hence, only the off-target
potential of the antisense strand need be considered
[0267] To identify potential off-target genes, the 21 mer sequences
corresponding to the candidate target regions plus a 3'-terminal AA
tail (to account for the TT overhangs) were subjected to a homology
search against publically available human mRNA sequences. To this
purpose, fastA (version 3.4) searches were performed with all 21mer
input sequences against a human RefSeq database (downloaded
available version from ftp://ftp.ncbi.nih.gov/refseq/ on
2005-07-25). fastA search was executed with
parameters-values-pairs--b 30-g 30 in order to take into account
the homology over the full length of the 21 mer. The search
resulted in a list of potential off-targets for candidate
siRNAs.
[0268] To sort the resulting list of potential off-targets, fastA
output files were analyzed to identify the host gene with the
highest off-target score. The following off-target properties for
each 21 mer input sequence were extracted for each potential
off-target to calculate the off-target score:
[0269] 1. Number of identical nucleotides to 21mer sequence
(Identity)
[0270] 2. Number of mismatches in seed region
[0271] The off-target score was calculated for considering
assumption 1 and 2 as follows: Identity-0.2*number of seed
mismatches
[0272] All siRNAs were sorted according to their highest off-target
score (ascending). An off-target score of 16.8 was used as a
cut-off for siRNA selection. 42 siRNAs specific for influenza A
matrix protein (MP), 3 siRNAs specific for influenza A nucleocapsid
protein (NP), and 1 siRNA specific for influenza A Polymerase Basic
protein 1 (PB1) had off target scores at or below this
threshold.
[0273] Given the comparatively low number of candidate siRNAs
resulting from the above selection procedure, the Positional
nucleotide numerical summary was re-examined with cut-off for
criterium 1 (target coverage) set to 70% and criterium 2 (target
specificity) remaining at 80%, followed by a repeat off-target
score ranking as described above. 2 additional siRNAs specific for
influenza A MP mRNA with a targeting efficiency of 79.9% were
additionally selected, for a total of 48 candidate siRNAs. The
sequences of these 48 candidate siRNAs are shown in Table 1A.
[0274] Because the selection process described above resulted only
in a limited number of candidate agents, the selection criteria
were somewhat relaxed to yield further candidate agents.
Specifically, criterium 1, above, was relaxed to 50% target
coverage, criterium 2, target efficiency, was kept at 80%, and the
above selection process was repeated. This procedure yielded the
additional agents AL-DP-8001 to AL-DP-8040, listed in Table 1C.
[0275] In this process, it was realized that the off-target scoring
step led to the greatest attrition rate in potential agents. In
order to obtain yet more candidate agents, the selection process
was therefore repeated once more, using criterium 1 at 80% target
coverage, criterium 2 at 80% target efficiency, and the off-target
scoring was omitted. This procedure yielded the additional agents
listed in Table 1D. Yet further candidate agents, listed in Table
1E, were obtained by repeating the selection once again, using
criterium 1 at 50% target coverage, criterium 2 at 80% target
efficiency, and omitting off-target scoring.
[0276] Additional candidate iRNA agents were identified by allowing
for the incorporation of universal bases. A Perl script was used to
first identify candidate sequences having target coverage and
target efficiency of 100% when the incorporation of up to 3
universal bases in the non-seed region (corresponding to positions
2-9 of the antisense strand) of the iRNA agent per strand. Table 1F
shows the agents identified in this manner. In a second round,
additional iRNA agents were identified that possess target coverage
and target efficiency of 80% when allowing for the incorporation of
one universal base. These iRNA agents are shown in Table 1G.
Example 2
siRNA Synthesis
[0277] Synthesis of Nucleotides Comprising Natural Bases
[0278] Single-stranded RNAs were produced by solid phase synthesis
on a scale of 1 .mu.mole using an Expedite 8909 synthesizer
(Applied Biosystems, Applera Deutschland GmbH, Darmstadt, Germany)
and controlled pore glass (CPG, 500 .ANG., Glen Research, Sterling
Va.) as solid support. RNA and RNA containing 2'-O-methyl
nucleotides were generated by solid phase synthesis employing the
corresponding phosphoramidites and 2'-O-methyl phosphoramidites,
respectively (Proligo Biochemie GmbH, Hamburg, Germany). These
building blocks were incorporated at selected sites within the
sequence of the oligoribonucleotide chain using standard nucleoside
phosphoramidite chemistry such as described in Current protocols in
nucleic acid chemistry, Beaucage, S. L. et al. (Edrs.), John Wiley
& Sons, Inc., New York, N.Y., USA. Phosphorothioate linkages
were introduced by replacement of the iodine oxidizer solution with
a solution of the Beaucage reagent (Chruachem Ltd, Glasgow, UK) in
acetonitrile (1%). Further ancillary reagents were obtained from
Mallinckrodt Baker (Griesheim, Germany).
[0279] Deprotection and purification by anion exchange HPLC of the
crude oligoribonucleotides were carried out according to
established procedures. Yields and concentrations were determined
by UV absorption of a solution of the respective RNA at a
wavelength of 260 nm using a spectral photometer (DU 640B, Beckman
Coulter GmbH, Unterschlei.beta.heim, Germany). Double stranded RNA
was generated by mixing an equimolar solution of complementary
strands in annealing buffer (20 mM sodium phosphate, pH 6.8; 100 mM
sodium chloride), heated in a water bath at 85-90.degree. C. for 3
minutes and cooled to room temperature over a period of 3-4 hours.
The purified RNA solution was stored at -20.degree. C. until
use.
[0280] As a result of the synthesis strategy described above, all
oligonucleotides synthesized as described above do not comprise a
phosphate group on their 5'-most nucleotide.
[0281] Synthesis of Nucleotides Comprising Universal Bases
Synthesis of Phosphoramidite and controlled pore glass support of
5'-O-(4,4'-dimethoxitrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-nitroind-
ole)-D-riboside
[0282] ##STR1##
Step A: 1-O-Methyl-D-riboside (102)
[0283] To a solution of D-ribose (25 g) in dry methanol (300 mL)
was added conc. sulfuric acid (1.88 mL) and stirred at room
temperature for 3 days. The reaction mixture was then neutralized
with 1 N sodium hydroxide solution and concentrated into a crude
residue. The crude residue was dissolved in methanol (200 mL) and
the solids were filtered off. The filtrate was concentrated into a
crude residue, which was applied to a column of silica gel eluted
with dichloromethane-methanol (5:1) to give a pure compound (23.0
g, 82%) as a syrup.
Step B: 1-O-Methyl-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside
(103)
[0284] To a solution of 1-O-methyl-D-riboside (13.43 g, 81.83
mmol), 18-crown-6 (1.34 g) in dry THF (100 mL) was added powdered
potassium hydroxide (69 g, 1.23 mol) and stirred at room
temperature for 40 to 60 min. 2,4-Dichlorobenzyl chloride (51 mL,
368.2 mmol) was added dropwise and the reaction mixture was stirred
at the same temperature overnight. The solids were filtered off and
the filtrate was concentrated into a crude residue which was
applied to a column of silica gel eluted with hexanes-ethyl acetate
(4:1) to give a pure compound (48 g, 92%) as a white solid.
[0285] .sup.1H-NMR (CDCl.sub.3, 400 MHz): .delta. 7.46-7.34 (m, 5H,
ArH), 7.24-7.16 (m, 4H, ArH), 4.99 (s, 1H, H-1), 4.71 (dd, 2H,
J.sub.gem=12.8 Hz, OCH.sub.2Ar), 4.63-4.61 (m, 4H, 2 OCH.sub.2Ar),
4.38-4.36 (m, 1H), 4.19-4.16 (dd, 1H), 3.98 (d, 1H, J=4.4 Hz), 3.75
(dd, 1H, J=3.6, J=10.2 Hz, H-5a), 3.66 (dd, 1H, J=3.6, J=10.4 Hz,
H-5b), 3.37 (s, 3H, OCH.sub.3).
Step B: 1-Bromo-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-ribose (104)
[0286] To a cold solution of
1-O-methyl-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside (3.22 g,
5.02 mmol) in dry dichloromethane (50 mL) cooled with ice-bath was
added HOAc-HBr (5.3 mL, 30%) and stirred at 0-25.degree. C. for 3
h. The reaction mixture was concentrated into a crude residue which
was co-evaporated with toluene (3.times.30 mL) into a crude residue
which was dried under a good vacuum and used for next reaction
without purification and identification as a syrup.
Step D:
1-(5-Nitroindole)-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside
(105)
[0287] To a solution of 5-nitroindole (2.44 g, 15.06 mmol) in dry
CH.sub.3CN (30 mL) was added sodium hydride (602 mg, 15.06 mmol,
60%) and stirred at room temperature for 3-4 h under an argon
atmosphere. The above obtained sugar donor (104) in dry CH.sub.3CN
(10 mL) was added and stirred at the same temperature under an
argon atmosphere overnight. The solids were filtered off and the
filtrate was concentrated into a crude residue which was applied to
a column of silica gel eluted with hexanes-ethyl acetate (3:1) to
give a pure compound 105 (2.16 g, 60%) as a .alpha. and .beta.
mixture (1:1).
Steps E, F:
5'-O-(4,4'-dimethoxitrityl)-1'-(5-nitroindole)-D-riboside (106) and
(107)
[0288] To a cold solution of
1-(5-nitroindole)-2,3,5-tri-O-(2,4-dichlorobenzyl)-D-riboside 105
(1.16 g, 1.51 mmol) in dry dichloromethane (100 mL) at -78.degree.
C. was added BCl.sub.3 in dichloromethane (23 mL, 1.0M) and stirred
at the same temperature for 2 h under an argon atmosphere and at
-40.degree. C. for 2 h. The reaction mixture was quenched with
methanol-dichloromethane (1:1, 50 mL) and neutralized with
ammonia-methanol solution. The solids were filtered off and the
filtrate was concentrated into a crude residue which was applied to
a column of silica gel eluted with dichloromethane-methanol (10:1)
to give a pure compound (300 mg, 68%) as a .alpha. and .beta.
mixture (1:1). To a solution of the above obtained compound (840
mg, 2.86 mmol) in dry pyridine (3-4 ml) and DMAP (90 mg) was added
DMTrCl (1.06 g) and stirred at room temperature under an argon
atmosphere overnight. The reaction mixture was concentrated into a
crude residue which was applied to a column of silica gel eluted
with hexanes-ethyl acetate (1:1) to give a pure compound 106 (550
mg) and compound 107 (190 mg), a mixture of compound 106 and 107
(360 mg).
[0289] Compound 106: .sup.1H-NMR (CDCl.sub.3, 2D g-COSY and 2D
NOESY, 400 MHz): .delta. 8.49 (d, 1H, J=1.6 Hz), 8.35 (d, 1H), 8.03
(dd, 1H, J=2.0, J=9.0 Hz), 7.70-7.69 (m, 2H), 7.47-7.14 (m, 8H,
ArH), 6.86-6.81 (m, 5H, ArH), 6.71 (d, 1H, J=3.6 Hz), 6.41 (d,
J=5.2 Hz, H'-1), 4.73 (t, 1H, J=4.8 Hz, H'-2), 4.46-4.42 (m, 3H,
H'-3, H'-4, H'-5), 3.79 (s, 6H, 2OCH.sub.3), 3.51 (dd, 1H, J=3.2,
J=10.4 Hz, H'-5a), 3.26 (dd, 1H, J=3.2, J=10.6 Hz, H'-5b).
[0290] Compound 107: .sup.1H-NMR (CDCl.sub.3, 2D g-COSY and 2D
NOESY, 400 MHz): .delta. 8.55 (d, 1H, J=2.0 Hz), 7.98 (dd, 1H,
J=2.4, J=9.2 Hz), 7.60 (d, 1H, J=9.2 Hz), 7.53 (d, 1H, J=3.2 Hz),
7.44-7.42 (m, 2H), 7.34-7.24 (m, 7H, ArH), 6.84-6.81 (m, 4H, ArH),
6.68 (d, 1H, J=3.2 Hz), 6.00 (d, 1H, J=5.2 Hz, H'-1), 4.53 (t, 1H,
J=7.6 Hz), 4.46-4.44 (m, 1H), 4.23-4.20 (m, 1H), 3.80-3.76 (m, 7H,
2OCH.sub.3, H'-5), 3.55 (dd, 1H, H'-5a), 3.43 (dd, 1H, H'-5b).
Step G:
5'-O-(4,4'-dimethoxitrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-n-
itroindole)-D-riboside (108) and
5'-O-(4,4'-dimethoxitrityl)-3'-O-(tert-butyldimethylsilyl)-1'-(5-nitroind-
ole)-D-riboside (109)
[0291] To a solution of
5'-O-(4,4'-dimethoxitrityl)-1'-(5-nitroindole)-D-riboside (106)
(550 mg, 0.92 mmol), AgNO.sub.3 (188 mg, 1.104 mmol), and pyridine
(0.74 mL, 9.2 mmol) in dry THF (9.2 mL) was added TBDMSCl (188 mg,
1.196 mmol) and stirred at room temperature under an argon
atmosphere overnight. The solids were filtered off and the filtrate
was concentrated into a crude residue which was applied to a column
of silica gel eluted with hexanes-ethyl acetate (4:1) to give a
pure compound 108 (230 mg, 35%), compound 109 (150 mg, 23%), and a
mixture of compound 108 and 109 (110 mg, 17%) in total yield of
75%.
[0292] Compound 108: .sup.1H-NMR (CDCl.sub.3, 2D g-COSY, 2D NOESY,
400 MHz): .delta. 8.56 (d, 1H, J=2.4 Hz), 7.88 (dd, 1H, J=2.4,
J=8.8 Hz), 7.62 (d, 1H, J=9.2 Hz), 7.54 (d, 1H, J=3.6 Hz),
7.46-7.44 (m, 2H), 7.36-7.25 (m, 6H, ArH), 6.85-6.83 (d, 5H, ArH),
6.69 (d, 1H, J=3.6 Hz), 5.94 (d, 1H, J=7.2 Hz, H'-1), 4.69 (dd, 1H,
H'-2), 4.31-4.29 (m, 2H, H'-3, H'-4), 3.80 (s, 6H, 2OCH.sub.3),
3.58 (dd, 1H, J=2.0, J=10.6 Hz, H'-5a), 3.40 (dd, 1H, J=2.0, J=10.4
Hz, H'-5b), 2.85 (d, 1H, J=0.8 Hz, 3'-OH), 0.78 (s, 9H, t-Bu),
-0.016 (s, 3H, SiCH.sub.3), -0.43 (s, 3H, SiCH.sub.3).
[0293] Compound 109: .sup.1H-NMR (CDCl.sub.3, 2D g-COSY, 2D NOESY,
400 MHz): .delta. 8.61 (d, 1H, J=2.4 Hz), 8.05 (dd, 1H, J=2.0,
J=8.8 Hz), 7.69-7.65 (m, 2H), 7.47-7.45 (m, 2H, ArH), 7.36-7.27 (m,
5H, ArH), 6.86-6.83 (m, 3H, ArH), 6.71 (d, 1H, J=3.2 Hz), 5.99 (d,
1H, J=4.8 Hz, H'-1), 4.51 (t, 1H, J=4.8 Hz, J=5.6 Hz, H'-3),
4.40-4.36 (m, 1H, H'-2), 4.17-4.15 (m, 2H, H'-4, H'-5), 3.82 (s,
3H, OCH.sub.3), 3.81 (s, 3H, OCH.sub.3), 3.63 (dd, 1H, J=2.4,
J=11.0 Hz, H'-5a), 3.31 (dd, 1H, J=2.8, J=11.0 Hz, H'-5b), 2.95 (d,
1H, J=6.0 Hz, 2'-OH), 0.91 (s, 9H, t-Bu), 0.05 (s, 3H, SiCH.sub.3),
0.00 (s, 3H, SiCH.sub.3).
Step H:
5'-O-(4,4'-dimethoxitrityl)-2'-O-(tert-butyldimethylsilyl)-1'-(5-n-
itroindole)-D-riboside-3'-O-cyanoethyl-N,N-diisopropylphosphoramidate
(110)
[0294] 2-Cyanoethyl-N,N-diisopropylchlorophosphoramidite (153 mg,
0.646 mmol) was added to a solution of
5'-O-(4,4'-dimethoxitrityl)-3'-O-(tert-butyldimethylsilyl)-1-(5-nitroindo-
le)-D-.beta.-riboside 108 (230 mg, 0.323 mmol),
diisopropylethylamine (306 uL, 1.78 mmol) and DMAP (10 mg) in dry
dichloromethane (3 mL) and stirred at room temperature for 4-6 h
under an argon atmosphere. The reaction mixture was concentrated to
a crude residue which was applied to a column of silica gel which
was saturated with 2% triethylamine in hexanes and eluted with
hexanes-ethyl acetate (2:1) to give a pure title compound 110 (250
mg, 85%) as an amorphous solid.
[0295] .sup.31P-NMR (CDCl3, 400 MHz): .delta. 149.54 (s), 146.57
(s). Anal. Cald of C.sub.50H.sub.65N.sub.4O.sub.9PSi: 924.43.
Found: 947.43 [M+Na].sup.+.
Step I: Solid supports of 2'-hydroxyl or 3'-hydroxyl of
5'-O-(4,4'-dimethoxitrityl)-1-(5-nitroindole)-D-riboside (111)
[0296] Succinic anhydride was added to a solution of a mixture of
2'-OTBDMS (108) or 3'-O-TBDMS of
5'-O-(4,4'-Dimethoxitrityl)-1-(5-nitroindole)-D-.beta.-riboside
(109), and DMAP in dry dichloromethane. The reaction mixture is
stirred at room temperature under an argon atmosphere for 6 h.
Another portion of succinct anhydrous and DMAP are added and
stirred for total of 16 h. The mixture is concentrated to a crude
residue which is dissolved in ethyl acetate (50 ml), washed with
citric acid (400 mg/20 ml), brine, and dried (Na.sub.2SO.sub.4).
The organic layer is concentrated to a crude nucleoside succinate
which was directly used for next reaction without further
purification.
[0297] Nucleoside succinate, DMAP, DTNP, and Ph.sub.3P are agitated
at room temperature for 20 min [Nucleoside and nucleotides, 1996,
15(4), 879-888.]. Then 1caa-CPG is added and agitated at the same
temperature for 45 min. The solids are filtered off and washed with
CH.sub.3CN, dichloromethane, and ether. The solid supports are
dried, capped under standard procedure, and washed to give solid
support.
[0298] The nitroindole-comprising Controlled Glass Support and
phosphoramidate thus obtained are employed in standard
oligonucleotide synthesis as described above for oligonucleotides
comprising natural bases.
Example 3
siRNA Testing In Vitro
[0299] The ability of the iRNA agents to inhibit replication of
influenza virus was tested in human cell lines in vitro, or is
tested in mice in vivo. The iRNA agent is transfected into the
cells, e.g., by transfection or electroporation, allowed to act on
the cells for a certain time, e.g., 24 hours, and levels of
infectivity were determined by a plaque forming or ELISA assay.
Complementing these direct assays, we tested the inhibition of
target gene expression by RNAi Agents for several influenza genes
recombinantly expressed in mammalian host cells.
[0300] Viruses and Cell Lines
[0301] Influenza virus A/PR/8/34 (PR8), subtype H1N1, was obtained
from Charles River Laboratories (ATCC #VR-1469). A/WSN/33 (WSN),
subtype H1N1, may be obtained from Thomas Chambers, University of
Kentucky, Lexington, Ky. USA (see Castrucci, M. R., et al., J.
Virol. 1992, 66:4647), or Dr. Peter Palese, Mount Sinai School of
Medicine New York City, N.Y., USA (see WO 04/028471). Virus stocks
were propagated in the allantoic cavity of embryonated hen eggs at
34.degree. C. for 48-72 h (PR8) or 37.degree. C. for 24 h (WSN)
(Tompkins, S. M., et al. Proc. Natl. Acad. Sci. 2004,
101:8682).
[0302] MDCK cells were obtained from the American Type Culture
Collection (ATCC, Rockville Md., USA; ATCC #CCL-34) and were grown
in MEM containing 8% heat-inactivated fetal bovine serum (FBS), 2
mM L-glutamine, 1 mM Sodium Pyruvate, 1.5 g/L sodium bicarbonate
and non-essential amino acids at 37.degree. C. under a 5%
CO.sub.2/95% air atmosphere.
[0303] Vero E6 African green monkey kidney epithelial cells were
obtained from ATCC (Rockville Md., USA, ATCC #CRL-1586) and were
grown in DMEM supplemented with 4.5 g/1 D-Glucose, 2 mM
L-Glutamine, 110 mg/l sodium pyruvate, 10% fetal bovine serum
(Hyclone, Cat #30070.03) and 0.1% Penicillin/Streptomycin at
37.degree. C. under a 5% CO.sub.2/95% air atmosphere.
[0304] Cos-7 African green monkey kidney cells were obtained from
the German Collection of Microorganisms and Cell Cultures (DSMZ,
Braunschweig, Germany, DSMZ #ACC 60) and were grown in Dulbecco's
MEM, 10% fetal calf serum, 2 mM L-glutamine, 1.2 .mu.g/ml sodium
bicarbonate, 100 u penicillin/100 .mu.g/ml streptomycin (Biochrom
AG, Berlin, Germany).
Example 3.1
Plaque Forming Assay
[0305] Cell Culture, siRNA Transfection, and Virus Infection.
[0306] MDCK cells were plated in 24-well plates at
7.5.times.10.sup.4 cells per well in 0.5 ml growth medium a day
before transfection. MDCK cells were 80% confluent the day of siRNA
transfection. Before transfection cells are fed with 0.25 ml growth
medium.
[0307] Prior to adding to cells, 1.5 ml (50 .mu.l per well) Optimem
I (Invitrogen) and 90 .mu.l (3 .mu.l per well) Lipofectamin 2000
(Invitrogen), the amount sufficient for transfection of one 24 well
plate, were combined in a 2 ml Sarstedt tube and incubated for
10-15 minutes at room temperature. The appropriate amount of siRNA
dissolved in annealing buffer is then added to the
Optimem/lipofectamine 2000 mixture to give the desired final
concentration, mixed, and incubated an additional 15-25 minutes at
room temperature. Next, 50 .mu.l of the siRNA/reagent complex is
added dropwise to each well as dictated by the experimental design.
Plates are then gently rocked to ensure complete mixing and
incubated at 37.degree. C. at 5% CO2/95% air for 14 hours.
[0308] Subsequently, the transfection medium was gently aspirated,
cells washed once with 0.25-0.5 ml of PBS, and 100 .mu.l of varying
concentrations of PR8 in MEM medium was added to each well. After
incubation at 37.degree. C. for 1-2 hour, 0.5 ml of overlay media
(MEM, 20 mM HEPES, 0.075% NaHCO3, 2 mM glutamine, 0.6% agarose, 0.5
.mu.g/ml TPCK-trypsin) were added, and plates incubated for 48 hrs
at 37.degree. C. in an incubator at 5% CO2/95% air. Plates were
then fixed and immunostained for viral plaques as described
below.
[0309] Immunostaining and Viral Quantitation
[0310] 48 hours post-infection, cells were fixed in neutral
buffered 10% formalin for 45 minutes, and wells rinsed with PBS.
Wells were then blocked with permeabilization buffer (1.times.PBS,
2% FBS, 0.5% saponin, 0.1% sodium azide) for 15 minutes at room
temperature, and 125 .mu.l of a solution containing 0.5 .mu.g/ml
mouse anti-influenza A biotinylated antibody MAB8258B (Chemicon)
was added. Following incubation for 1 hr at room temperature, wells
were rinsed twice with PBS to remove unbound antibody, and 125
.mu.l of a solution of 1 .mu.g/ml of horse radish peroxidase (HRP)
conjugated streptavidin (Vector Laboratories) in PBS per well was
added, plates incubated for 45 min, and washed three times with
PBS. 200 .mu.l of TMB substrate (Vector Laboratories #SK-4400) per
well were added. Following incubation for 5-10 minutes at room
temperature in the dark, the calorimetric reaction was stopped with
distilled water, the water discarded and the plates air-dried.
Stained influenza plaques were counted by inverted light microscopy
at 4.times. magnification. Plaque forming activity was compared to
cells transfected with Lipofectamin only (mock-treated), and
expressed in terms of [(plaque forming activity in treated
cells)/(plaque forming activity in mock-treated cells)].times.100=%
remaining infectivity
Example 3.2
ELISA Assay
[0311] MDCK or Vero cells were plated in 96-well plates at 10.sup.4
cells per well in 0.1 ml growth medium a day before transfection.
The cells were 80% confluent the day of siRNA transfection. Before
transfection, cells were fed with 44 .mu.l growth medium.
[0312] 1.08 ml (9 .mu.l per well) Optimem I (Invitrogen) and 42
.mu.l (0.35 .mu.l per well) Lipofectamin 2000 (Invitrogen), the
amount sufficient for transfection of one 96 well plate, were
combined in a 2 ml Sarstedt tube and incubated for 10-15 minutes at
room temperature. The appropriate amount of siRNA dissolved in
annealing buffer was then added to the Optimem/lipofectamine 2000
mixture to give the desired final concentration, mixed, and
incubated an additional 15-25 minutes at room temperature. Next, 10
.mu.l of the siRNA/reagent complex was added to each well as
dictated by the experimental design. Plates were gently rocked to
ensure complete mixing and then incubated at 37.degree. C. in an
incubator at 5% CO.sub.2/95% air for 14 hours.
[0313] Subsequently, cells were washed once with PBS, infected with
PR8 influenza virus in 50 .mu.l of MEM per well, and incubated for
1-2 hours. Thereafter, plates were washed once with PBS, and 200
.mu.l of MEM with 0.25/0.5 .mu.g/ml (MDCK/VERO, respectively) of
trypsin were added. Two days post infection, plates were fixed in
10% Buffered Formalin for 15 min. Cells were rinsed with PBS,
blocked with blocking buffer for 15 min. at RT, and 50 .mu.l of a
solution containing 0.5 .mu.g/ml of biotinylated anti-influenza A
monoclonal antibody MAB8258B (Chemicon) per well were added. Plates
were incubated at RT for 1 hour, washed twice with PBS, and 50
.mu.l per well of a solution containing 1 .mu.g/ml of AP-conjugated
streptavidin (Vector Laboratories) in blocking buffer was added.
After incubation for 45 min and washing 3.times. with PBS, 100
.mu.l per well of pNPP substrate solution was added. Plates were
developed at RT in the dark and read at 405 nm.
Example 3.3
Inhibition of Recombinantly Expressed Influenza Target Genes by
siRNA
[0314] Consensus sequences of MP (SEQ ID NO: 1453), NP (SEQ ID NO:
1454), PA (SEQ ID NO: 1455), PB1 (SEQ ID NO: 1456) and PB2 (SEQ ID
NO: 1457) (see Table 5) were synthesized by GENEART (Regensburg,
Germany) and cloned into GENEART standard vectors. MP and PA were
subcloned into psiCheck-2 (Promega, Mannheim, Germany) via AsiSI
and NotI (both NEBn, Frankfurt, Germany) sites, NP, (PB1) and PB2
via XhoI and NotI, resulting in a construct with the flu gene
between the stop-codon and the polyA-signal of Renilla luciferase.
Correct cloning was confirmed by end sequencing performed by GATC
Biotech (Konstanz, Germany).
[0315] Transfections:
[0316] Cos-7 cells were seeded at 1.5.times.10.sup.4 cells/well on
white 96-well plates with clear bottom (Greiner Bio-One GmbH,
Frickenhausen, Germany) in 75 .mu.l of growth medium. Directly
after seeding the cells, 50 ng of plasmid/well were transfected
with Lipofectamine2000 (Invitrogen) as described below for the
siRNAs, with the plasmid diluted in Opti-MEM to a final volume of
12.5 .mu.l/well, prepared as a mastermix for the whole plate.
[0317] siRNA transfections were performed in quadruplicates 4 h
after plasmid transfection. For each well 0.5 .mu.l
Lipofectamine2000 (Invitrogen GmbH, Karlsruhe, Germany) were mixed
with 12 .mu.l Opti-MEM (Invitrogen) and incubated for 15 min at
room temperature. For an siRNA concentration of 50 nM in the 100
.mu.l transfection volume, 1 .mu.l of a 5 .mu.M siRNA were mixed
with 11.5 .mu.l Opti-MEM per well, combined with the
Lipofectamine2000-Opti-MEM mixture and again incubated for 15
minutes at room temperature. During incubation, the growth medium
was removed from cells and replaced by 75 .mu.l/well of fresh
medium. siRNA-Lipofectamine2000-complexes were applied completely
(25 .mu.l each per well) to the cells and cells were incubated for
24 h at 37.degree. C. and 5% CO.sub.2 in a humidified incubator
(Heraeus GmbH, Hanau, Germany).
[0318] Cells were harvested by removing growth medium and
application of 150 .mu.l of a 1:1 mixture consisting of medium and
Dual-Glo Luciferase substrate, from the Dual-Glo Luciferase Assay
System (Promega, Mannheim, Germany). The luciferase assay was
performed according to the manufacturer's protocol for Dual-Glo
Luciferase assay and luminescence was measured in a Victor-Light
1420 Luminescence Counter (Perkin Elmer, Rodgau-Jugesheim,
Germany). Values obtained with Renilla luciferase were normalized
to the respective values obtained with Firefly luciferase. Values
acquired with siRNAs directed against an influenza gene were
normalized to the value obtained with an unspecific siRNA (directed
against neomycin resistance gene) set to 100%.
[0319] Effective siRNAs from the screen were further characterized
by dose response curves. Transfections of dose response curves were
performed at the following siRNA concentrations according to the
above protocol: 100 nM, 25 nM, 6.3 nM, 1.6 nM, 400 pM, 100 pM, 24
pM, 6 pM, 1.5 pM, 380 fM. IC.sub.50 values determined by
parametrized curve fitting using the program XLfit. TABLE-US-00013
TABLE 5 Virtual consensus sequences for influenza genes MP (SEQ ID
NO: 1453), NP (SEQ ID NO: 1454), PA (SEQ ID NO: 1455), PB1 (SEQ ID
NO: 1456) and PB2 (SEQ ID NO: 1457) for cloning into Cos-7 cells
MP: virtual consensus derived from gi|13383290|gb|AB049165|
Influenza A virus (A/parakeet/Chiba/1/97(H9N2)) M1, M2 genes for
membrane ion channel, matrix protein, complete ads. SEQ ID NO: 1453
ATGAGTCTTC TAACCGAGGT CGAAACGTAC GTTCTCTCTA TCATCCCGTC AGGCCCCCTC
60 AAAGCCGAGA TCGCGCAGAG ACTTGAAGAT GTCTTTGCAG AGAAGAACAC
AGATCTCGAG 120 GCTCTCATGG AATGGCTAAA GACAAGACCA ATCCTGTCAC
CTCTGACTAA GGGGATTTTA 180 GGGTTTGTGT TCACGCTCAC CGTGCCCAGT
GAGCGAGGAC TGCAGCGTAG ACGCTTTGTC 240 CAGAATGCCC TAAATGGGAA
TGGAGACCCA AACAACATGG ACAGGGCAGT TAAACTATAC 300 AAGAAGCTGA
AGAGGGAAAT AACATTCCAT GGGGCTAAGG AAGTTGCACT CAGTTACTCT 360
GCTGGTGCAC TTGCCAGTTG CATGGGTCTC ATATACAACC GGATGGGAAC AGTGACCACA
420 GAAGTGGCTC TTGGCCTAGT GTGTGCCACT TGTGAGCAGA TTGCAGATTC
ACAACATCGG 480 TCCCACAGGC AGATGGCGAC TACCACCAAC CCACTAATCA
GACATGAGAA CAGAATGGTG 540 CTGGCCAGCA CTACAGCTAA GGCTATGGAG
CAGATGGCTG GATCAAGTGA GCAGGCAGCG 600 GAAGCCATGG AAGTCGCAAG
TCAGGCTAGG CAGATGGTGC AGGCAATGAG GACAATTGGG 660 ACTCATCCTA
GCTCCAGTGC AGGTCTAAAA GATAATCTTC TTGAAAATTT GCAGGCCTAC 720
CAGAAACGAA TGGGGGTGCA GATGCAGCGA TTCAAGTGAT CCTCTCGTTG TTGCAGCAAG
780 TATCATTGGG ATCTTGCACT TGATATTGTG GATTCTTGAT CGTCTTTTCT
TCAAATGCAT 840 TTATCGTCGC CTTAAATACG GTTTGAAAAG AGGGCCTTCT
ACGGAAGGAG TACCTGAGTC 900 TATGAGGGAA GAGTATCGAC AGGAACAGCA
GAGTGCTGTG GATGTTGACG ATGGTCATTT 960 TGTCAACATA GAGCTGGAGT AA 982
NP: virtual consensus derived from H5N1 gi|14326108|AF370122|
Influenza A virus (A/Goose/Guangdong/3/97(H5N1)) nucleoprotein
gene, complete cds. SEQ ID NO: 1454 CTCACTGAGT GACATCAAAA
TCATGGCGTC TCAAGGCACC AAACGATCTT ATGAACAGAT 60 GGAAACTGGT
GGAGAACGCC AGAATGCTAC TGAGATCAGA GCATCTGTTG GAAGAATGGT 120
TGGTGGAGTT GGGAGGTTTT ATATACAGAT GTGCACTGAA CTCAAACTCA GCGACTATGA
180 AGGAAGGCTG ATTCAGAACA GCATAACAAT AGAGAGAATG GTTGTCTCTG
CATTTGATGA 240 AAGGAGGAAC AAATACCTGG AAGAACATCC CAGTGCGGGG
AAGGACCCAA AGAAAACTGG 300 AGGTCCAATC TACCGAAGAA GAGACGGGAA
ATGGGTGAGA GAGCTGATTC TGTATGACAA 360 AGAGGAGATC AGGAGAATTT
GGCGTCAAGC GAACAATGGA GAAGATGCAA CTGCTGGTCT 420 CACTCACCTG
ATGATCTGGC ATTCCAATCT AAATGATGCC ACATACCAGA GAACAAGAGC 480
TCTCGTGCGT ACTGGGATGG ACCCTAGAAT GTGCTCTCTG ATGCAAGGAT CAACTCTCCC
540 GAGGAGATCT GGAGCTGCTG GTGCGGCAGT AAAGGGAGTC GGAACTATGG
TGATGGAACT 600 AATTCGGATG ATAAAGCGAG GGATTAACGA TCGGAATTTC
TGGAGAGGTG AAAATGGGCG 660 AAGAACAAGG ATTGCATATG AGAGAATGTG
CAACATTCTC AAAGGGAAAT TCCAAACAGC 720 AGCACAAAGA GCAATGATGG
ATCAGGTACG GGAAAGCAGA AATCCTGGGA ATGCTGAGAT 780 CGAAGATCTC
ATATTTCTGG CACGGTCTGC ACTCATCCTG AGAGGATCAG TGGCCCACAA 840
GTCCTGCTTG CCTGCTTGTG TGTACGGGCT TGCCGTGGCC AGTGGATATG ACTTTGAGAG
900 AGAAGGGTAC TCTCTGGTCG GGATTGATCC TTTCCGTCTG CTGCAAAACA
GCCAGGTCTT 960 TAGTCTAATT AGACCAAATG AGAATCCAGC ACATAAAAGT
CAATTGGTGT GGATGGCATG 1020 CCATTCTGCA GCATTTGAAG ATCTGAGAGT
CTCAAGCTTC ATCAGAGGGA CAAGAGTGGC 1080 CCCAAGGGGA CAACTATCTA
CTAGAGGAGT ACAAATTGCT TCAAATGAGA ACATGGAAAC 1140 AATGGACTCC
AGCACTCTTG AACTGAGAAG CAGATATTGG GCTATAAGGA CCAGGAGTGG 1200
AGGAAACACC AACCAGCAGA GAGCATCTGC AGGACAAATC AGTGTGCAGC CTACTTTCTC
1260 GGTACAGAGA AATCTTCCCT TCGAAAGAGC GACCATTATG GCGGCATTCA
CAGGGAATAC 1320 AGAGGGCAGA ACATCTGACA TGAGGACTGA AATCATAAGG
ATGATGGAAA GCTCCAGACC 1380 AGAAGATGTG TCTTTCCAGG GGCGGGGAGT
CTTCGAGCTC TCGGACGAAA AGGCAACGAA 1440 CCCGATCGTG CCTTCCTTTG
ACATGAGTAA TGAAGGATCT TATTTCTTCG GAGACAATGC 1500 AGAGGAGTAT
GACAATTGAA G 1521 PA: virtual consensus derived from
H5N1gi|47156500|AY585473| Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) polymerase (PA) mRNA, complete ads.
SEQ ID NO: 1455 ATGGAAGACT TTGTGCGACA ATGCTTCAAT CCAATGATTG
TCGAGCTTGC GGAAAAGGCA 60 ATGAAAGAAT ATGGGGAAGA TCCGAAAATC
GAAACGAACA AATTTGCAGC AATATGCACA 120 CACTTAGAAG TCTGTTTCAT
GTATTCAGAT TTTCACTTTA TTGATGAACG GGGCGAATCA 180 ATAATTGTAG
AATCTGGCGA TCCGAATGCA TTATTGAAAC ACCGATTTGA AATAATTGAA 240
GGAAGAGACC GAACAATGGC CTGGACAGTG GTGAATAGTA TCTGCAACAC CACAGGAGTT
300 GAGAAACCTA AATTTCTCCC AGATTTGTAT GACTACAAAG AGAACCGATT
CATTGAAATT 360 GGAGTGACAC GGAGGGAAGT TCATATATAC TATCTAGAGA
AAGCCAACAA GATAAAATCC 420 GAGAAGACAC ACATTCACAT ATTCTCATTC
ACTGGGGAGG AAATGGCCAC CAAAGCGGAC 480 TACACCCTTG ATGAAGAGAG
CAGGGCAAGA ATCAAAACCA GGCTGTTCAC CATAAGGCAG 540 GAAATGGCCA
GTAGGGGTCT ATGGGATTCC TTTCGTCAGT CCGAGAGAGG CGAAGAGACA 600
ATTGAAGAAA GATTTGAAAT CACAGGAACC ATGCGCAGGC TTGCCGACCA AAGTCTCCCA
660 CCGAACTTCT CCAGCCTTGA AAACTTTAGA GCCTATGTGG ATGGATTCGA
ACCGAACGGC 720 TGCATTGAGG GCAAGCTTTC TCAAATGTCA AAAGAAGTGA
ACGCCAGAAT TGAGCCATTT 780 CTGAAGACAA CACCACGCCC TCTCAGATTA
CCTGATGGGC CTCCCTGCTC TCAGCGGTCG 840 AAGTTCTTGC TGATGGATGC
CCTTAAATTA AGCATCGAAG ACCCGAGTCA TGAGGGGGAG 900 GGGATACCGC
TATATGATGC AATCAAATGC ATGAAAACAT TTTTCGGCTG GAAAGAGCCC 960
AACATCGTAA AACCACATGA AAAAGGCATA AACCCCAATT ACCTCCTGGC TTGGAAGCAA
1020 GTGCTGGCAG AACTCCAAGA TATTGAAAAT GAGGAGAAAA TCCCAAAAAC
AAAGAACATG 1080 AAGAAAACAA GCCAATTGAA GTGGGCACTC GGTGAGAACA
TGGCACCAGA GAAAGTAGAC 1140 TTTGAGGATT GCAAAGATGT TAGCGATCTA
AGACAGTATG ACAGTGATGA ACCAGAGCCT 1200 AGATCACTAG CAAGCTGGAT
CCAGAGTGAA TTCAACAAGG CATGTGAATT GACAGATTCG 1260 AGTTGGATTG
AACTTGATGA AATAGGGGAA GACGTTGCTC CAATTGAGCA CATTGCAAGT 1320
ATGAGAAGGA ACTATTTCAC AGCGGAAGTA TCCCATTGCA GGGCCACTGA ATACATAATG
1380 AAGGGGGTGT ACATAAACAC AGCTCTGTTG AATGCATCCT GTGCAGCCAT
GGATGACTTT 1440 CAACTGATTC CAATGATAAG CAAATGCAGA ACCAAAGAAG
GAAGACGGAA AACTAACCTG 1500 TATGGATTCA TTATAAAAGG AAGATCCCAT
TTGAGGAATG ATACCGATGT GGTAAACTTT 1560 GTGAGTATGG AATTCTCTCT
TACTGACCCG AGGCTGGAGC CACACAAGTG GGAAAAGTAC 1620 TGTGTTCTCG
AGATAGGAGA CATGCTCCTA CGGACTGCAA TAGGCCAAGT TTCAAGGCCC 1680
ATGTTCCTGT ATGTGAGAAC CAATGGAACC TCCAAGATCA AAATGAAATG GGGAATGGAG
1740 ATGAGGCGAT GCCTTCTTCA ATCCCTTCAA CAGATTGAGA GCATGATTGA
GGCCGAGTCT 1800 TCTGTCAAAG AGAAAGACAT GACCAAAGAA TTCTTTGAAA
ACAAATCAGA AACATGGCCA 1860 ATTGGAGAGT CACCCAAAGG AGTGGAGGAA
GGCTCCATCG GGAAGGTGTG CAGAACCTTA 1920 CTGGCGAAAT CTGTGTTCAA
CAGTCTATAT GCATCTCCAC AACTCGAGGG GTTTTCAGCT 1980 GAATCAAGAA
AATTGCTTCT CATTGTTCAG GCACTTAGGG ACAACCTGGA ACCTGGGACC 2040
TTCGATCTTG GAGGGCTATA TGAAGCAATT GAGGAGTGCC TGATTAATGA TCCCTGGGTT
2100 TTTGCTTAATG CGTCTTGGTT CAACTCCTTC CTCACACATG CACTGAAATA GTT
2153 PB1: virtual consensus derived from H5N1gi|58531084|AB166860|
Influenza A virus (A/chicken/Yamaguchi/7/2004(H5N1)) PB1 gene for
polymerase basic protein 1, complete ads. SEQ ID NO: 1456
ATGGATGTCA ATCCGACTTT ACTTTTCTTG AAAGTACCAG TGCAAAATGC TATAAGTACC
60 ACATTCCCTT ATACTGGAGA CCCTCCATAC AGCCATGGAA CAGGGACAGG
ATACACCATG 120 GACACAGTCA ACAGAACACA CCAATATTCA GAAAAGGGGA
AGTGGACAAC AAACACAGAG 180 ACTGGAGCAC CCCAACTCAA CCCGATTGAT
GGACCACTAC CTGAGGATAA TGAGCCCTGT 240 GGGTATGCAC AAACAGATTG
TGTATTGGAA GCAATGGCTT TCCTTGAAGA ATCCCACCCA 300 GGGATCTTTG
AAAACTCGTG TCTTGAAACG ATGGAAATTG TTCAACAAAC AAGAGTGGAT 360
AAACTGACCC AAGGTCGCCA GACCTATGAC TGGACATTGA ATAGAAACCA ACCGGCTGCA
420 ACTGCTTTGG CCAACACTAT AGAAATCTTC AGATCGAACG GTCTAACAGC
CAATGAATCG 480 GGACGGCTAA TAGATTTCCT CAAGGATGTG ATGGAATCAA
TGGATAAGGA AGAAATGGAG 540 ATAACAACAC ATTTCCAGAG AAAGAGAAGA
GTGAGGGACA ACATGACCAA GAAAATGGTC 600 ACACAAAGAA CAATAGGGAA
GAAAAAACAA AGGCTGAACA AAAAGAGCTA CCTGATAAGA 660 GCACTGACAC
TGAACACAAT GACAAAAGAT GCAGAAAGAG GCAAATTGAA GAGGCGAGCA 720
ATTGCAACAC CCGGAATGCA AATCAGAGGA TTCGTGTACT TTGTTGAAAC ACTAGCGAGG
780 AGTATCTGTG AGAAACTTGA GCAATCTGGA CTCCCAGTCG GAGGGAATGA
GAAGAAGGCT 840 AAATTGGCAA ACGTCGTGAG GAAGATGATG ACTAACTCAC
AAGATACTGA ACTCTCCTTT 900 ACAATTACTG GAGACAATAC CAAATGGAAT
GAGAATCAGA ATCCTAGGAT GTTTCTGGCA 960 ATGATAACGT ACATCACAAG
GAACCAGCCA GAATGGTTTC GGAATGTCTT AAGCATTGCC 1020 CCTATAATGT
TCTCAAACAA AATGGCGAGA TTAGGAAAAG GATACATGTT CGAAAGTAAG 1080
AGCATGAAGT TACGAACACA AATACCAGCA GAAATGCTTG CAAACATTGA TCTCAAATAC
1140 TTCAATGAAT TAACGAAAAA GAAAATTGAG AAAATAAGAC CTCTATTAAT
AGATGGTACA 1200 GCCTCATTGA GCCCTGGAAT GATGATGGGC ATGTTCAACA
TGCTGAGTAC AGTCCTAGGA 1260 GTCTCAATCC TGAATCTTGG ACAGAAAAGG
TACACCAAAA CCACATATTG GTGGGACGGA 1320 CTCCAATCCT CTGATGATTT
CGCTCTCATC GTAAATGCAC CGAATCATGA GGGAATACAA 1380 GCAGGAGTGG
ATAGGTTTTA TAGGACTTGT AAACTAGTTG GAATCAATAT GAGCAAGAAG 1440
AAGTCTTACA TAAATCGGAC AGGGACATTT GAATTCACGA GCTTTTTCTA CCGCTATGGA
1500 TTTGTAGCCA ATTTCAGTAT GGAGCTGCCC AGTTTTGGAG TGTCTGGAAT
TAATGAATCG 1560 GCCGACATGA GCATTGGTGT TACAGTGATA AAGAACAATA
TGATAAACAA CGACCTTGGG 1620 CCAGCAACAG CTCAGATGGC TCTTCAGCTA
TTCATCAAGG ACTACAGATA CACATACCGA 1680 TGCCACAGAG GGGATACGCA
AATCCAAACG AGGAGATCAT TCGAGCTGAA GAAGCTGTGG 1740 GAGCAAACCC
GTTCAAAGGC AGGACTGTTG GTTTCAGATG GAGGACCAAA TCTATACAAT 1800
ATCCGAAATC TCCATATTCC TGAGGTCTGC TTAAAATGGG AATTGATGGA TGAAGATTAC
1860 CAGGGCAGAC TGTGTAATCC TCTGAATCCG TTCGTCAGCC ATAAGGAAAT
TGAATCTGTC 1920 AACAATGCTG TAGTAATGCC AGCTCATGGC CCGGCCAAAA
GCGTGGAATA TGATGCCGTT 1980 GCAACTACAC ATTCATGGAT TCCTAAAAGG
AATCGTTCCA TTCTCAATAC GAGTCAAAGG 2040 GGAATTCTTG AGGATGAACA
GATGTACCAG AAGTGCTGCA ATCTATTCGA GAAATTCTTC 2100 CCCAGCAGTT
CATATCGGAG GCCAGTTGGA ATTTCCAGCA TGGTGGAGGC CATGGTGTCT 2160
AGGGCCCGAA TTGACGCACG AATTGATTTC GAGTCTGGAA GGATTAAGAA AGAAGAGTTT
2220 GCTGAGATCA TGAAGATCTG TTCCACCATT GAAGAGCTCA GACGGCAAAA ATAG
2274 PB2: virtual consensus derived from H5N1gi|19697859|AY059525|
Influenza A virus (A/Duck/Hong Kong/2986.1/2000(H5N1)) segment 1
polymerase (PB2) gene, partial cds. SEQ ID NO: 1457 ATGGAGAGAA
TAAAAGAATT AAGAGATCTA ATGTCGCAGT CTCGCACTCG CGAGATACTA 60
ACAAAAACCA CTGTGGACCA TATGGCCATA ATCAAGAAAT ACACATCAGG AAGACAAGAG
120 AAGAACCCTG CTCTCAGAAT GAAATGGATG ATGGCAATGA AATATCCAAT
CACAGCAGAC 180 AAGAGAATAA TAGAGATGAT TCCTGAAAGG AATGAACAAG
GGCAGACGCT TTGGAGCAAG 240 ACAAATGATG CTGGATCGGA CAGGGTGATG
GTGTCTCCCC TAGCTGTAAC TTGGTGGAAT 300 AGGAATGGGC CGACGACAAG
TGCAGTCTAT TATCCAAAGG TTTACAAAAC ATACTTTGAG 360 AAGGTTGAAA
GGTTAAAACA TGGAACCTTC GGTCCCGTTC ATTTCCGAAA CCAAATTAAA 420
ATACGCCGCC GAGTTGATAT AAATCCTGGC CATGCAGATC TCAATGCTAA AGAAGCACAA
480 GATGTCATCA TGGAGGTCGT TTTCCCAAAT GAAGTGGGAG CTAGAATATT
GACATCAGAG 540 TCGCAATTGA CAATAACGAA AGAAAAGAAA GAAGAGCTCC
AGGATTGTAA GATTGCTCCT 600 TTAATGGTTG CATACATGTT GGAAAGGGAA
CTGGTCCGCA AAACCAGATT CCTACCGGTA 660 GCAGGCGGAA CAAGCAGTGT
GTACATTGAG GTATTGCATT TGACTCAAGG GACCTGCTGG 720 GAACAGATGT
ACACTCCAGG CGGAGAAGTG AGAAATGACG ATGTTATCCA GAGTATGATC 780
ATCGCTGCCA GAAACATTGT TAGGAGAGCA ACGGTATCAG CGGATCCACT GGCATCACTG
840 CTGGAGATGT GTCACAGCAC ACAAATTGGT GGGATAAGGA TGGTGGACAT
CCTTAGGCAA 900 AATCCAACTG AGGAACAAGC TGTGGATATA TGCAAAGCAG
CAATGGGTTT GAGGATCAGT 960 TCATCCTTTA GCTTTGGAGG CTTCACTTTC
AAAAGAACAA GTGGAACATC CGTCAAGAAG 1020 GAAGAGGAAG TGCTTACAGG
CAACCTCCAA ACATTGAAAA TAAGAGTACA TGAGGGGTAT 1080 GAGGAATTCA
CAATGGTTGG GCGGAGGGCA ACAGCTATCC TGAGGAAAGC AACTAGAAGG 1140
CTGATTCAGT TGATAGTAAG TGGAAGAGAC GAACAATCAA TCGCTGAGGC AATCATTGTA
1200 GCAATGGTGT TCTCACAGGA GGATTGCATG ATAAAGGCAG TCCGAGGCGA
TCTGAATTTC 1260 GTAAACAGAG CAAACCAAAG ATTAAACCCC ATGCATCAAC
TCCTGAGACA TTTTCAAAAG 1320 GATGCAAAAG TGCTATTTCA GAATTGGGGA
ATTGAACCCA TTGATAATGT CATGGGGATG 1380 ATCGGAATAT TACCTGACAT
GACTCCCAGC ACAGAAATGT CACTGAGAGG AGTAAGAGTT 1440 AGTAAAATGG
GAGTGGATGA ATATTCCAGC ACTGAGAGAG TAGTTGTAAG TATTGACCGT 1500
TTCTTAAGGG TTCGAGATCA GCGGGGGAAC GTACTCTTAT CTCCCGAAGA GGTCAGCGAA
1560 ACACAGGGAA CAGAGAAATT GGCAATAACA TATTCATCAT CAATGATGTG
GGAAATCAAC 1620 GGTCCTGAGT CAGTGCTTGT TAACACCTAT CAATGGATCA
TCAGAAACTG GGAGACTGTG 1680 AAGATTCAAT GGTCTCAAGA CCCCACGATG
CTGTACAATA AGATGGAGTT TGAACCGTTC 1740 CAATCCTTGG TACCTAAAGC
TGCCAGAGGT CAATACAGTG GATTTGTGAG AACACTATTC 1800 CAACAAATGC
GTGACGTACT GGGGACATTT GATACTGTCC AGATAATAAA GCTGCTACCA 1860
TTTGCAGCAG CCCCACCGGA GCAGAGCAGA ATGCAGTTTT CTTCTCTAAC TGTGAATGTG
1920 AGAGGCTCAG GAATGAGAAT ACTTGTAAGG GGCAATTCCC CTGTGTTCAA
CTACAATAAG 1980 GCAACCAAAA GGCTTACCGT TCTTGGAAAG GACGCAGGTG
CATTAACAGA GGATCCAGAT 2040 GAGGGAACAG CCGGAGTGGA ATCTGCAGTA
CTGAGGGGAT TCCTAATTCT AGGCAAGGAG 2100 GACAAAAGAT ATGGACCAGC
ATTGAGCATC AATGAACTGA GCAATCTTGC GAAAGGGGAG 2160 AAAGCTAATG
TGCTGATAGG GCAAGGAGAC GTGGTGTTGG TAATGAAACG GAAACGGGAC 2220
TCTAGCATAC TTACTGACAG CCAGACAGCG ACCAAAAGAA TTCGGATGGC CATCAATTAG
2280
[0320] Table 1A, C, D and E list the duplex identifier, the
sequences of sense and antisense strand, the agents' target genes,
and the results from the above assays, where performed, for
selected exemplary agents of the invention. Table 1B and H list the
duplex identifier, the duplex identifier of the corresponding
unmodified sequence, the sequences of sense and antisense strand,
and the agents' target genes, for selected exemplary agents bearing
modified nucleic acids groups, in order to stabilize these agents
against degradation, in which all pyrimidine base comprising
nucleotides comprised a 2'-O-methyl group in the sense strand, and
all pyrimidine base comprising nucleotides in a sequence context of
5'-ca-3' or 5'-ua-3' comprised a 2'-O-methyl group in the antisense
strand, except for those agents where the antisense strand does not
comprise nucleotides in a sequence context of 5'-ca-3' or 5'-ua-3',
in which all uridines in a sequence context of 5'-ug-3' are
2'-O-methyl-modified nucleotides in the antisense strand (e.g.
AL-DP-2295, AL-DP-2301, and AL-DP-2302). Table 2 lists
concentrations at 50% maximal inhibition calculated from the dose
response determinations in Cos-7 cells engineered to express
influenza genes for some particularly preferred RNAi agents of the
invention. TABLE-US-00014 TABLE 6 Sequences used in analysis of
Influenza A Matrix Protein (MP) AY180470 Influenza A virus strain
A/Quail/Nanchang/12-340/2000 (H1N1) matrix protein (M) gene,
partial cds. AY633213 Influenza A virus (A/mallard/Alberta/211/98
(H1N1)) matrix protein (M) gene, complete cds. AY664487 Influenza A
virus (A/mallard/Alberta/119/98 (H1N1)) nonfunctional matrix
protein mRNA, partial sequence. M55476 Influenza virus type A
matrix protein (M1) gene, complete cds and M2 protein (M2) gene,
complete cds. M55479 Influenza virus type A matrix protein (M1)
gene, complete cds and M2 protein (M2) gene, complete cds. M55480
Influenza virus type A matrix protein (M1) gene, complete cds and
M2 protein (M2) gene, complete cds. M63528 Influenza A virus
(A/turkey/Minnesota/166/81 (H1N1)) membrane protein M1 and membrane
protein M2 genes, complete cds. U49119 Influenza A virus matrix
proteins M1 and M2 (M) gene, complete cds. Z26859 Influenza virus
type A M and M2 genes for matrix proteins Z26860 Influenza virus
type A M and M2 genes for matrix proteins AY422021 Influenza A
virus (A/duck/Hokkaido/95/01(H2N2)) matrix protein 1 (M) gene,
partial cds. M12699 Avian Influenza A/Mallard/NY/6750/78 RNA
segment 7 encoding M1 and M2 proteins, complete cds. AF213915
Influenza A virus (A/Chicken/Italy/5945/95(H3N2)) segment 7 matrix
protein (M) gene, partial cds. AY180498 Influenza A virus strain
A/Chicken/Nanchang/3-120/2001 (H3N2) matrix protein (M) gene,
partial cds. AY664458 Influenza A virus (A/ruddy
turnstone/Delaware/142/99 (H3N2)) nonfunctional matrix protein
mRNA, partial sequence. AY769614 Influenza A virus
(A/turkey/Ohio/313053/04(H3N2)) matrix protein gene, partial cds.
AY779257 Influenza A virus (A/turkey/North Carolina/12344/03(H3N2))
matrix protein 2 (M) gene, partial cds; and matrix protein 1 (M)
gene, complete cds. AY779258 Influenza A virus
(A/turkey/Minnesota/764-2/03(H3N2)) matrix protein 2 (M) gene,
partial cds; and matrix protein 1 (M) gene, complete cds. AY862623
Influenza A virus (A/chicken/Korea/S6/03(H3N2)) matrix protein (M)
gene, complete cds. AY862624 Influenza A virus
(A/duck/Korea/S7/03(H3N2)) matrix protein (M) gene, complete cds.
AY862625 Influenza A virus (A/duck/Korea/S8/03(H3N2)) matrix
protein (M) gene, complete cds. AY862626 Influenza A virus
(A/duck/Korea/S9/03(H3N2)) matrix protein (M) gene, complete cds.
AY862627 Influenza A virus (A/duck/Korea/S10/03(H3N2)) matrix
protein (M) gene, complete cds. AY862628 Influenza A virus
(A/dove/Korea/S11/03(H3N2)) matrix protein (M) gene, complete cds.
Z26858 Influenza virus type A M and M2 genes for matrix proteins
AB166865 Influenza A virus (A/chicken/Yamaguchi/7/2004(H5N1)) M1
and M2 genes for matrix protein and membrane ion channel, complete
cds. AB188819 Influenza A virus (A/chicken/Oita/8/2004(H5N1)) M2,
M1 genes for membrane ion channel 2, matrix protein 1, complete
cds. AF509043 Influenza A virus (A/Chicken/Hong Kong/FY150/01
(H5N1)) M1 protein (M1) gene, complete cds. AF509044 Influenza A
virus (A/Pheasant/Hong Kong/FY155/01 (H5N1)) M1 protein (M1) gene,
complete cds. AF509045 Influenza A virus (A/Silky Chicken/Hong
Kong/SF189/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509046
Influenza A virus (A/Quail/Hong Kong/SF203/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509047 Influenza A virus (A/Pigeon/Hong
Kong/SF215/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509048
Influenza A virus (A/Chicken/Hong Kong/SF219/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509049 Influenza A virus (A/Chicken/Hong
Kong/715.5/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509050
Influenza A virus (A/Chicken/Hong Kong/751.1/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509051 Influenza A virus (A/Chicken/Hong
Kong/822.1/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509052
Influenza A virus (A/Chicken/Hong Kong/829.2/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509053 Influenza A virus (A/Chicken/Hong
Kong/830.2/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509054
Influenza A virus (A/Chicken/Hong Kong/858.3/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509055 Influenza A virus (A/Chicken/Hong
Kong/866.3/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509056
Influenza A virus (A/Chicken/Hong Kong/867.1/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509057 Influenza A virus (A/Chicken/Hong
Kong/879.1/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509058
Influenza A virus (A/Chicken/Hong Kong/873.3/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509059 Influenza A virus (A/Chicken/Hong
Kong/876.1/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509060
Influenza A virus (A/Chicken/Hong Kong/891.1/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509061 Influenza A virus (A/Chicken/Hong
Kong/893.2/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509062
Influenza A virus (A/Goose/Hong Kong/76.1/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509063 Influenza A virus (A/Goose/Hong
Kong/ww100/01 (H5N1)) M1 protein (M1) gene, complete cds. AF509064
Influenza A virus (A/Duck/Hong Kong/573.4/01 (H5N1)) M1 protein
(M1) gene, complete cds. AF509065 Influenza A virus (A/Duck/Hong
Kong/646.3/01 (H5N1)) M1 protein (M1) gene, complete cds. AY059506
Influenza A virus (A/Goose/Hong Kong/ww26/2000(H5N1)) segment 7
matrix protein (M) gene, partial cds. AY059507 Influenza A virus
(A/Goose/Hong Kong/ww28/2000(H5N1)) segment 7 matrix protein (M)
gene, partial cds. AY059508 Influenza A virus (A/Duck/Hong
Kong/ww381/2000(H5N1)) segment 7 matrix protein (M) gene, partial
cds. AY059509 Influenza A virus (A/Duck/Hong Kong/ww461/2000(H5N1))
segment 7 matrix protein (M) gene, partial cds. AY059510 Influenza
A virus (A/Goose/Hong Kong/ww491/2000(H5N1)) segment 7 matrix
protein (M) gene, partial cds. AY059511 Influenza A virus
(A/Duck/Hong Kong/2986.1/2000(H5N1)) segment 7 matrix protein (M)
gene, partial cds. AY059512 Influenza A virus (A/Goose/Hong
Kong/3014.8/2000(H5N1)) segment 7 matrix protein (M) gene, partial
cds. AY075029 Influenza A virus (A/Chicken/Hong
Kong/317.5/2001(H5N1)) matrix protein 1 and matrix protein 2 (M)
gene, complete cds. AY075035 Influenza A virus (A/Duck/Hong
Kong/380.5/2001(H5N1)) matrix protein 1 and matrix protein 2 (M)
gene, complete cds. AY221530 Influenza A virus
(A/Chicken/HongKong/NT873.3/01-MB(H5N1)) matrix protein (M) gene,
complete cds. AY221531 Influenza A virus
(A/Chicken/HongKong/NT873.3/01(H5N1)) matrix protein (M) gene,
complete cds. AY221532 Influenza A virus
(A/Chicken/HongKong/FY150/01-MB(H5N1)) matrix protein (M) gene,
complete cds. AY221533 Influenza A virus
(A/Chicken/HongKong/FY150/01(H5N1)) matrix protein (M) gene,
complete cds. AY221534 Influenza A virus
(A/Pheasant/HongKong/FY155/01-MB(H5N1)) matrix protein (M) gene,
complete cds. AY221535 Influenza A virus
(A/Pheasant/HongKong/FY155/01(H5N1)) matrix protein (M) gene,
complete cds. AY221536 Influenza A virus
(A/Chicken/HongKong/YU822.2/01-MB(H5N1)) matrix protein (M) gene,
complete cds. AY221537 Influenza A virus
(A/Chicken/HongKong/YU822.2/01(H5N1)) matrix protein (M) gene,
complete cds. AY221538 Influenza A virus
(A/Chicken/HongKong/YU562/01(H5N1)) matrix protein (M) gene,
complete cds. AY518361 Influenza A virus
(A/duck/China/E319-2/03(H5N1)) membrane ion channel M2 and matrix
protein M1 (M) gene, complete cds. AY575895 Influenza A virus
(A/Gs/HK/739.2/02 (H5N1)) matrix protein (M) gene, complete cds.
AY575896 Influenza A virus (A/Eg/HK/757.3/02 (H5N1)) matrix protein
(M) gene, partial cds. AY575897 Influenza A virus
(A/G.H/HK/793.1/02 (H5N1)) matrix protein (M) gene, partial cds.
AY575898 Influenza A virus (A/Dk/HK/821/02 (H5N1)) matrix protein
(M) gene, partial cds. AY575899 Influenza A virus (A/Ck/HK/31.4/02
(H5N1)) matrix protein (M) gene, complete cds. AY575900 Influenza A
virus (A/Ck/HK/61.9/02 (H5N1)) matrix protein (M) gene, complete
cds. AY575901 Influenza A virus (A/Ck/HK/YU777/02 (H5N1)) matrix
protein (M) gene, complete cds. AY575902 Influenza A virus
(A/Ck/HK/96.1/02 (H5N1)) matrix protein (M) gene, complete cds.
AY575903 Influenza A virus (A/Ck/HK/409.1/02 (H5N1)) matrix protein
(M) gene, complete cds. AY575904 Influenza A virus
(A/Ph/HK/sv674.15/02 (H5N1)) matrix protein (M) gene, complete cds.
AY585378 Influenza A virus (A/duck/Fujian/01/2002(H5N1)) matrix
protein mRNA, complete cds. AY585379 Influenza A virus
(A/duck/Fujian/13/2002(H5N1)) matrix protein mRNA, complete cds.
AY585380 Influenza A virus (A/duck/Fujian/17/2001(H5N1)) matrix
protein mRNA, complete cds. AY585381 Influenza A virus
(A/duck/Fujian/19/2000(H5N1)) matrix protein mRNA, complete cds.
AY585382 Influenza A virus (A/duck/Guangdong/01/2001(H5N1)) matrix
protein mRNA, complete cds. AY585383 Influenza A virus
(A/duck/Guangdong/07/2000(H5N1)) matrix protein mRNA, complete cds.
AY585384 Influenza A virus (A/duck/Guangdong/12/2000(H5N1)) matrix
protein mRNA, complete cds. AY585385 Influenza A virus
(A/duck/Guangdong/22/2002(H5N1)) matrix protein mRNA, complete cds.
AY585386 Influenza A virus (A/duck/Guangdong/40/2000(H5N1)) matrix
protein mRNA, complete cds. AY585387 Influenza A virus
(A/duck/Guangxi/07/1999(H5N1)) matrix protein mRNA, complete cds.
AY585388 Influenza A virus (A/duck/Guangxi/22/2001(H5N1)) matrix
protein mRNA, partial cds. AY585389 Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) matrix protein mRNA, complete cds.
AY585390 Influenza A virus (A/duck/Guangxi/53/2002(H5N1)) matrix
protein mRNA, complete cds. AY585391 Influenza A virus
(A/duck/Shanghai/08/2001(H5N1)) matrix protein mRNA, complete cds.
AY585392 Influenza A virus (A/duck/Shanghai/13/2001(H5N1)) matrix
protein mRNA, complete cds. AY585393 Influenza A virus
(A/duck/Shanghai/35/2002(H5N1)) matrix protein mRNA, complete cds.
AY585394 Influenza A virus (A/duck/Shanghai/37/2002(H5N1)) matrix
protein mRNA, complete cds. AY585395 Influenza A virus
(A/duck/Shanghai/38/2001(H5N1)) matrix protein mRNA, complete cds.
AY585396 Influenza A virus (A/duck/Zhejiang/11/2000(H5N1)) matrix
protein mRNA, complete cds. AY585397 Influenza A virus
(A/duck/Zhejiang/52/2000(H5N1)) matrix protein mRNA, complete cds.
AY585398 Influenza A virus (A/duck/Guangxi/50/2001(H5N1)) matrix
protein mRNA, complete cds. AY590578 Influenza A virus
(A/chicken/Nakorn-Patom/Thailand/CU- K2/2004(H5N1)) matrix protein
M2 and matrix protein M1 (M) gene, partial and complete cds.
AY609315 Influenza A virus (A/chicken/Guangdong/174/04(H5N1))
segment 7, complete sequence. AY651374 Influenza A virus
(A/Ck/Indonesia/BL/2003(H5N1)) membrane ion channel 2 (M) gene,
partial cds; and matrix protein 1 (M) gene, complete cds. AY651375
Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651376
Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651377
Influenza A virus (A/Ck/Indonesia/2A/2003(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651378
Influenza A virus (A/Ck/Indonesia/4/2004(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651379
Influenza A virus (A/Ck/Indonesia/5/2004(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651380
Influenza A virus (A/Ck/Thailand/1/2004(H5N1)) membrane ion channel
2 and matrix protein 1 (M) gene, complete cds. AY651381 Influenza A
virus (A/Ck/Thailand/73/2004(H5N1)) membrane ion channel 2 (M)
gene, partial cds; and matrix protein 1 (M) gene, complete cds.
AY651382 Influenza A virus (A/Ck/Thailand/9.1/2004(H5N1)) membrane
ion channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651383 Influenza A virus
(A/Qa/Thailand/57/2004(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds.
AY651384 Influenza A virus (A/bird/Thailand/3.1/2004(H5N1))
membrane ion channel 2 (M) gene, partial cds; and matrix protein 1
(M) gene, complete cds. AY651385 Influenza A virus
(A/Dk/Thailand/71.1/2004(H5N1)) membrane ion channel 2 (M) gene,
partial cds; and matrix protein 1 (M) gene, complete cds. AY651386
Influenza A virus (A/Gs/Thailand/79/2004(H5N1)) membrane ion
channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651391 Influenza A virus (A/Ck/Viet
Nam/33/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M)
gene, complete cds. AY651392 Influenza A virus (A/Ck/Viet
Nam/35/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M)
gene, complete cds. AY651393 Influenza A virus (A/Ck/Viet
Nam/36/2004(H5N1)) membrane ion channel 2 (M) gene, partial cds;
and matrix protein 1 (M) gene, complete cds. AY651394 Influenza A
virus (A/Ck/Viet Nam/37/2004(H5N1)) membrane ion channel 2 and
matrix protein 1 (M) gene, complete cds. AY651395 Influenza A virus
(A/Ck/Viet Nam/38/2004(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds. AY651396 Influenza A virus
(A/Ck/Viet Nam/39/2004(H5N1)) membrane ion channel 2 (M) gene,
partial cds; and matrix protein 1 (M) gene, complete cds. AY651397
Influenza A virus (A/Ck/Viet Nam/C57/2004 (H5N1)) membrane ion
channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651398 Influenza A virus (A/Dk/Viet
Nam/11/2004(H5N1)) membrane ion channel 2 and matrix protein 1 (M)
gene, complete cds. AY651399 Influenza A virus (A/Gf/HK/38/2002
(H5N1)) membrane ion channel 2 and matrix protein 1 (M) gene,
partial cds. AY651400 Influenza A virus (A/Ck/HK/31.2/2002(H5N1))
membrane ion channel 2 and matrix protein 1 (M) gene, complete cds.
AY651401 Influenza A virus (A/Ck/HK/37.4/2002(H5N1)) membrane ion
channel 2 and matrix protein 1 (M) gene, complete cds. AY651402
Influenza A virus (A/SCk/HK/YU100/2002(H5N1)) membrane ion channel
2 and matrix protein 1 (M) gene, complete cds. AY651403 Influenza A
virus (A/Ck/HK/YU22/2002(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds. AY651404 Influenza A virus
(A/Ck/HK/3176.3/2002(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, partial cds. AY651405 Influenza A virus
(A/Ck/HK/3169.1/2002(H5N1)) matrix protein 1 and membrane ion
channel 2 (M) gene, partial cds. AY651406 Influenza A virus
(A/Ck/HK/FY157/2003(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds. AY651407 Influenza A virus
(A/Ck/HK/YU324/2003(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds. AY651408 Influenza A virus
(A/Ck/HK/2133.1/2003(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, partial cds. AY651409 Influenza A virus
(A/Ck/HK/NT93/2003(H5N1)) membrane ion channel 2 (M) gene, partial
cds; and matrix protein 1 (M) gene, complete cds. AY651410
Influenza A virus (A/Ck/HK/SSP141/2003(H5N1)) membrane ion channel
2 (M) gene, partial cds; and matrix protein 1 (M) gene, complete
cds. AY651411 Influenza A virus (A/Ck/HK/WF157/2003(H5N1)) membrane
ion channel 2 and matrix protein 1 (M) gene, complete cds. AY651412
Influenza A virus (A/peregrine falcon/HK/D0028/2004(H5N1)) membrane
ion channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651413 Influenza A virus (A/black headed
gull/HK/12.1/2003(H5N1)) membrane ion channel 2 and matrix protein
1 (M) gene, complete cds. AY651414 Influenza A virus (A/grey
heron/HK/861.1/2002(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, complete cds. AY651415 Influenza A virus
(A/feral pigeon/HK/862.7/2002(H5N1)) membrane ion channel 2 and
matrix protein 1 (M) gene, complete cds. AY651416 Influenza A virus
(A/tree sparrow/HK/864/2002(H5N1)) matrix protein 1 and membrane
ion channel 2 (M) gene, partial cds. AY651417 Influenza A virus
(A/teal/China/2978.1/2002(H5N1)) membrane ion channel 2 and matrix
protein 1 (M) gene, partial cds. AY651418 Influenza A virus
(A/Dk/HN/5806/2003(H5N1)) membrane ion channel 2 (M) gene, partial
cds; and matrix protein 1 (M) gene, complete cds. AY651419
Influenza A virus (A/Dk/ST/4003/2003(H5N1)) membrane ion channel 2
(M) gene, partial cds; and matrix protein 1 (M) gene, complete cds.
AY651420 Influenza A virus (A/Ck/ST/4231/2003(H5N1)) membrane ion
channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651421 Influenza A virus (A/Dk/YN/6255/2003(H5N1))
membrane ion channel 2 (M) gene, partial cds; and matrix protein 1
(M) gene, complete cds. AY651422 Influenza A virus
(A/Dk/YN/6445/2003(H5N1)) membrane ion channel 2 (M) gene, partial
cds; and matrix protein 1 (M) gene, complete cds. AY651423
Influenza A virus (A/Ck/YN/374/2004(H5N1)) membrane ion channel 2
(M) gene, partial cds; and matrix protein 1 (M) gene, complete cds.
AY651424 Influenza A virus (A/Dk/HN/101/2004(H5N1)) membrane ion
channel 2 (M) gene, partial cds; and matrix protein 1 (M) gene,
complete cds. AY651425 Influenza A virus (A/Dk/HN/303/2004(H5N1))
membrane ion channel 2 (M) gene, partial cds; and matrix protein 1
(M) gene, complete cds. AY651426 Influenza A virus
(A/Ph/ST/44/2004(H5N1)) membrane ion channel 2 (M) gene, partial
cds; and matrix protein 1 (M) gene, complete cds. AY651427
Influenza A virus (A/Ck/YN/115/2004(H5N1)) membrane ion channel 2
(M) gene, partial ads; and matrix protein 1 (M) gene, complete cds.
AY653194 Influenza A virus (A/chicken/Jilin/9/2004(H5N1)) segment
7, complete sequence. AY676045 Influenza A virus strain
(A/duck/Hong Kong/821/02(H5N1)) membrane protein (M) gene, complete
cds. AY676046 Influenza A virus strain (A/egret/Hong
Kong/757.2/03(H5N1)) membrane protein (M) gene, complete cds.
AY676047 Influenza A virus strain (A/chicken/Korea/ES/03(H5N1))
membrane protein (M) gene, complete cds. AY676048 Influenza A virus
strain (A/duck/Korea/ESD1/03(H5N1)) membrane protein (M) gene,
complete cds. AY684709 Influenza A virus
(A/chicken/Hubei/327/2004(H5N1)) matrix protein 2 (M2) and matrix
protein 1 (M1) genes, complete cds. AY737292 Influenza A virus
(A/chicken/Guangdong/191/04(H5N1)) segment 7, complete sequence.
AY737298 Influenza A virus (A/chicken/Guangdong/178/04(H5N1))
segment 7, complete sequence. AY737306 Influenza A virus
(A/duck/Guangdong/173/04(H5N1)) segment 7, complete sequence.
AY770077 Influenza A virus (A/chicken/Hubei/489/2004(H5N1)) matrix
protein 2 (M2) and matrix protein 1 (M1) genes, complete cds.
AY770998 Influenza A virus (A/chicken/Ayutthaya/Thailand/CU-
23/04(H5N1)) matrix protein gene, complete cds. AY818145 Influenza
A virus (A/chicken/Vietnam/C58/04(H5N1)) matrix protein M1 gene,
complete cds. AY818146 Influenza A virus
(A/quail/Vietnam/36/04(H5N1)) matrix protein M1 gene, complete cds.
AY856865 Influenza A virus (A/duck/Shandong/093/2004(H5N1)) segment
7, complete sequence. DQ055851 Influenza A virus
(A/chicken/Yunnan/K001/2004(H5N1)) matrix protein M1 gene, complete
cds. AB189048 Influenza A virus (A/chicken/Kyoto/3/2004(H5N1)) M2,
M1 genes for membrane ion channel; M2, matrix protein 1, complete
cds,. AB189056 Influenza A virus (A/crow/Kyoto/53/2004(H5N1)) M2,
M1 genes for membrane ion channel; M2, matrix protein 1, complete
cds,. AB189064 Influenza A virus (A/crow/Osaka/102/2004(H5N1)) M2,
M1 genes for membrane ion channel; M2, matrix protein 1, complete
cds,. AF046082 Influenza A virus (A/Chicken/Hong Kong/220/97
(H5N1)) matrix protein 2 (M2) and matrix protein 1 (M1) genes,
complete cds. AF098560 Influenza A virus (A/Chicken/Hong
Kong/258/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein (M)
genes, partial cds. AF098561 Influenza A virus (A/Chicken/Hong
Kong/y388/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein
(M) genes, partial cds. AF098562 Influenza A virus (A/Chicken/Hong
Kong/728/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein (M)
genes, partial cds. AF098563 Influenza A virus (A/Chicken/Hong
Kong/786/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein (M)
genes, partial cds. AF098564 Influenza A virus (A/Chicken/Hong
Kong/915/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein (M)
genes, partial cds. AF098566 Influenza A virus (A/Duck/Hong
Kong/p46/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein (M)
genes, partial cds. AF098567 Influenza A virus (A/Duck/Hong
Kong/y283/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein
(M) genes, partial cds. AF098568 Influenza A virus (A/Goose/Hong
Kong/w355/97 (H5N1)) M1 matrix protein (M) and M2 matrix protein
(M) genes, partial cds. AF144306 Influenza A virus
(A/Goose/Guangdong/1/96(H5N1)) matrix proteins M1 and M2 (M) gene,
alternatively spliced products, complete cds. AF216711 Influenza A
virus (A/Environment/Hong Kong/437-4/99 (H5N1)) matrix protein 1
and matrix protein 2 genes, complete cds. AF216719 Influenza A
virus (A/Environment/Hong Kong/437-6/99 (H5N1)) matrix protein 1
and matrix protein genes, complete cds. AF216727 Influenza A virus
(A/Environment/Hong Kong/437-8/99 (H5N1)) matrix protein 1 and
matrix protein 2 genes, complete cds. AF216735 Influenza A virus
(A/Environment/Hong Kong/437-10/99 (H5N1)) matrix protein 1 and
matrix protein 2 genes, complete cds. AF359560 Influenza A virus
(A/Goose/Guangdong/3/97(H5N1)) matrix protein 1 and matrix protein
2 (M) gene, complete cds. AF398429 Influenza A virus (A/Goose/Hong
Kong/385.3/2000(H5N1)) matrix protein 1 (M) gene, partial cds.
AF398430 Influenza A virus (A/Goose/Hong Kong/385.5/2000(H5N1))
matrix protein 1 (M) gene, partial cds. AF468843 Influenza A virus
(A/Duak/Anyang/AVL-1/2001(H5N1)) matrix protein 1 and matrix
protein 2 genes, complete cds. AF509040 Influenza A virus
(A/Chicken/Hong Kong/FY77/01 (H5N1)) M1 protein (M1) gene, complete
cds. AF509041 Influenza A virus (A/Chicken/Hong Kong/YU562/01
(H5N1)) M1 protein (M1) gene, complete cds. AF509042 Influenza A
virus (A/Chicken/Hong Kong/YU563/01 (H5N1)) M1 protein (M1) gene,
complete cds. AF073180 Influenza A virus (A/Chicken/New
Jersey/15086-3/94 (H7N3NSA)) matrix protein 1 (M1) and matrix
protein 2 (M2) genes, complete cds. AF073197 Influenza A virus
(A/Turkey/Oregon/71 (H7N3NSB)) matrix protein 1 (M1) and matrix
protein 2 (M2) genes, complete cds. AY664433 Influenza A Virus
(A/ruddy turnstone/New Jersey/65/85(H7N3)) nonfunctional matrix
protein mRNA, partial sequence. AY677732 Influenza A virus
(A/chicken/British Columbia/CN7-3/04 (H7N3)) matrix protein 1 (M1)
gene, complete cds. AF073198 Influenza A virus
(A/Turkey/Colorado/13356/91 (H7N3NSA)) matrix protein 1 (M1) and
matrix protein 2 (M2) genes, complete cds. AF073200 Influenza A
virus (A/Quail/Arkansas/16309-7/94(H7N3NSA)) matrix protein 1 (M1)
and matrix protein 2 (M2) genes, complete cds. AF073201 Influenza A
virus (A/Turkey/Utah/24721-10/95 (H7N3NSA)) matrix protein 1 (M1)
and matrix protein 2 (M2) genes, complete cds. AJ627492 Influenza A
virus (A/turkey/Italy/214845/2002(H7N3)) gene for membrane protein
1 and gene for membrane protein 2, genomic RNA. AJ627497 Influenza
A virus (A/turkey/Italy/220158/2002(H7N3)) gene for membrane
protein 1 and gene for membrane protein 2, genomic RNA. AY241600
Influenza A virus (A/Chicken/New York/12273-11/99(H7N3)) matrix
protein 1 and matrix protein 2 genes, complete cds. AY241602
Influenza A virus (A/chicken/NY/14714-9/99(H7N3)) matrix protein 1
and matrix protein 2 genes, complete cds. AY241615 Influenza A
virus (A/Duck/NJ/117228-7/01(H7N3)) matrix protein 1 and matrix
protein 2 genes, complete cds. AY241616 Influenza A virus
(A/Duck/PA/143585/01(H7N3)) matrix protein 1 and matrix protein 2
genes, complete cds. AY300975 Influenza A virus (A/Blue-winged
Teal/TX/2/01 (H7N3) membrane protein (M) gene, complete cds.
AY303652 Influenza A virus (A/chicken/Chile/176822/02(H7N3)) matrix
protein 1 and matrix protein 2 genes, complete cds. AY303653
Influenza A virus (A/chicken/Chile/4322/02(H7N3)) matrix protein 1
and matrix protein 2 genes, complete cds. AY303654 Influenza A
virus (A/chicken/Chile/4957/02(H7N3)) matrix protein 1 gene,
complete cds; and matrix protein 2 gene, partial cds. AY303655
Influenza A virus (A/chicken/Chile/4968/02(H7N3)) matrix protein 1
and matrix protein 2 genes, complete cds. AY303656 Influenza A
virus (A/chicken/Chile/4977/02(H7N3)) matrix protein 1 and matrix
protein 2 genes, complete cds. AY303657 Influenza A virus
(A/turkey/Chile/4418/02(H7N3)) matrix protein 1 gene, complete cds;
and matrix protein 2 gene, partial cds. AY586427 Influenza A Virus
(A/turkey/Italy/214845/02 (H7N3)) matrix protein gene, partial cds.
AY586428 Influenza A virus (A/turkey/Italy/220158/2002 (H7N3))
matrix protein gene, partial cds. AY586429 Influenza A Virus
(A/mallard/Italy/43/01(H7N3)) matrix protein gene, partial cds.
AY586430 Influenza A virus (A/mallard/Italy/33/01(H7N3)) matrix
protein gene, partial cds. AY611525 Influenza A virus
(A/chicken/British Columbia/04(H7N3)) matrix protein 2 (M) and
matrix protein 1 (M) genes, complete cds. AY646079 Influenza A
virus (A/chicken/British Columbia/GSC_human_B/04(H7N3)) matrix
protein 2 and matrix protein 1 (M) gene, complete cds. AY648288
Influenza A virus (A/GSC_chicken_B/British Columbia/04(H7N3))
matrix protein 2 (M) and matrix protein 1 (M) genes, complete cds.
AY650271 Influenza A virus (A/GSC_chicken/British
Columbia/04(H7N3)) matrix protein 2 (M) and matrix protein 1 (M)
genes, complete cds. AJ619676 Influenza A virus
(A/chicken/Germany/R28/03(H7N7)) M1 gene for membrane protein 1,
genomic RNA. AY340086 Influenza A virus
(A/Netherlands/124/03(H7N7)) matrix protein gene, partial cds.
AY340087 Influenza A virus (A/Netherlands/126/03(H7N7)) matrix
protein gene, partial cds. AY340088 Influenza A virus
(A/Netherlands/127/03(H7N7)) matrix protein gene, partial cds.
AY340089 Influenza A virus (A/Netherlands/219/03(H7N7)) matrix
protein gene, complete cds. AY340090 Influenza A virus
(A/Netherlands/33/03(H7N7)) matrix protein gene, complete cds.
AY340091 Influenza A virus (A/chicken/Netherlands/1/03(H7N7))
matrix protein gene, complete cds. AY664468 Influenza A virus
(A/ruddy turnstone/Delaware/134/99 (H7N7)) nonfunational matrix
protein mRNA, partial sequenae. L37795 Influenza virus
A/chicken/Brescia/1902 (H7N7) matrix protein
(M1) gene and transmembrane protein (M2) gene, complete cds. L37796
Influenza virus A/FPV/Dobson (H7N7) matrix protein (M1) gene and
transmembrane protein (M2) gene, complete cds. L37797 Influenza
virus A/FPV/Weybridge (H7N7) matrix protein (M1) gene and
transmembrane protein (M2) gene, complete cds. M23917 Influenza
A/chicken/FPV/Weybridge (H7N7) M1 matrix protein gene, complete
cds. M23921 Influenza A/chicken/FPV/Weybridge (H7N7) M2 matrix
protein gene, complete cds. M38299 Influenza A/FPV/Weybridge (H7N7)
matrix (M) protein (seg 7) gene, complete cds. M63523 Influenza A
virus (A/chicken/Victoria/1/85 (H7N7)) membrane protein M1 and
membrane protein M2 genes, complete cds. M63526 Influenza virus
type A (strain A/FPV/Dobson/27 (H7N7)) membrane protein M1 and
membrane protein M2 genes, complete cds. AB049165 Influenza A virus
(A/parakeet/Chiba/1/97(H9N2)) M1, M2 genes for membrane ion
channel, matrix protein, complete cds. AB049166 Influenza A virus
(A/parakeet/Narita/92A/98(H9N2)) M1, M2 genes for membrane ion
channel, matrix protein, complete cds. AF222671 Influenza A virus
(A/Silky Chicken/Hong Kong/SF44/99(H9N2)) segment 7 M1 (M1) gene,
partial cds. AF508684 Influenza A virus (A/Ostrich/South
Africa/9508103/95(H9N2)) segment 7 matrix protein M1 (M) gene,
complete cds. AF508685 Influenza A virus
(A/Chicken/Pakistan/4/99(H9N2)) segment 7 matrix protein M1 (M)
gene, partial cds. AF508686 Influenza A virus
(A/Chicken/Pakistan/5/99(H9N2)) segment 7 matrix protein M1 (M)
gene, partial cds. AF508687 Influenza A virus
(A/Chicken/Germany/R45/98 (H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508688 Influenza A virus
(A/Duck/Germany/113/95(H9N2)) segment 7 matrix protein M1 (M) gene,
complete cds. AF508689 Influenza A virus
(A/Chicken/Iran/11T/99(H9N2)) segment 7 matrix protein M1 (M) gene,
partial cds. AF508690 Influenza A virus (A/Chicken/Saudi
Arabia/532/99(H9N2)) segment 7 matrix protein M1 (M) gene, partial
cds. AF508691 Influenza A virus (A/Pheasant/Ireland/PV18/97(H9N2))
segment 7 matrix protein M1 (M) gene, complete cds. AF508692
Influenza A virus (A/Chicken/Korea/99029/99(H9N2)) segment 7 matrix
protein M1 (M) gene, complete cds. AF508693 Influenza A virus
(A/Chicken/Beijing/8/98(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508694 Influenza A virus
(A/Chicken/Guangdong/10/00(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508695 Influenza A virus
(A/Chicken/Guangdong/11/97(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508696 Influenza A virus
(A/Chicken/Heilongjiang/10/97(H9N2)) segment 7 matrix protein M1
(M) gene, complete cds. AF508697 Influenza A virus
(A/Chicken/Henan/62/00(H9N2)) segment 7 matrix protein M1 (M) gene,
complete cds. AF508698 Influenza A virus
(A/Chicken/Ningxia/5/99(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508699 Influenza A virus
(A/Chicken/Sichuan/5/97(H9N2)) segment 7 matrix protein M1 (M)
gene, partial cds. AF508700 Influenza A virus
(A/Chicken/Shandong/6/96(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508701 Influenza A virus
(A/Chicken/Shijiazhuang/2/99(H9N2)) segment 7 matrix protein M1 (M)
gene, partial cds. AF508702 Influenza A virus
(A/Chicken/Shenzhen/9/97(H9N2)) segment 7 matrix protein M1 (M)
gene, complete cds. AF508703 Influenza A virus
(A/Duck/Nanjing/1/97(H9N2)) segment 7 matrix protein M1 (M) gene,
complete cds. AF508704 Influenza A virus
(A/Quail/Shanghai/8/96(H9N2)) segment 7 matrix protein M1 (M) gene,
complete cds. AF523482 Influenza A virus
(A/Duck/Shantou/1043/00(H9N2)) matrix protein (M) gene, complete
cds. AF523483 Influenza A virus (A/Duck/Shantou/2134/00(H9N2))
matrix protein (M) gene, complete cds. AF523484 Influenza A virus
(A/Wild Duck/Shantou/4808/01(H9N2)) matrix protein (M) gene,
complete cds. AF523485 Influenza A virus
(A/Duck/Shantou/1042/00(H9N2)) matrix protein (M) gene, complete
cds. AF523486 Influenza A virus (A/Duck/Shantou/2143/00(H9N2))
matrix protein (M) gene, complete cds. AF523487 Influenza A virus
(A/Duck/Shantou/2144/00(H9N2)) matrix protein (M) gene, complete
cds. AF523488 Influenza A virus (A/Duck/Shantou/1881/00(H9N2))
matrix protein (M) gene, complete cds. AF523489 Influenza A virus
(A/Duck/Shantou/1796/00(H9N2)) matrix protein (M) gene, complete
cds. AF523490 Influenza A virus (A/Duck/Shantou/2102/00(H9N2))
matrix protein (M) gene, complete cds. AF523491 Influenza A virus
(A/Duck/Shantou/830/00(H9N2)) matrix protein (M) gene, complete
cds. AF523492 Influenza A virus (A/Duck/Shantou/2088/01(H9N2))
matrix protein (M) gene, complete cds. AF523493 Influenza A virus
(A/Duck/Shantou/1605/01(H9N2)) matrix protein (M) gene, complete
cds. AF523494 Influenza A virus (A/Duck/Hong Kong/610/79(H9N2))
matrix protein (M) gene, complete cds. AF523495 Influenza A virus
(A/Duck/Hong Kong/552/79(H9N2)) matrix protein (M) gene, complete
cds. AF523496 Influenza A virus (A/Duck/Hong Kong/289/78(H9N2))
matrix protein (M) gene, complete cds. AF523497 Influenza A virus
(A/Duck/Hong Kong/86/76(H9N2)) matrix protein (M) gene, complete
cds. AF523498 Influenza A virus (A/Duck/Hong Kong/366/78(H9N2))
matrix protein (M) gene, partial cds. AF536719 Influenza A virus
(A/Chicken/Beijing/1/95(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536720 Influenza A virus
(A/Chicken/Beijing/2/97(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536721 Influenza A virus
(A/Chicken/Beijing/3/99(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536722 Influenza A virus
(A/Chicken/Guangdong/97(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536723 Influenza A virus
(A/Chicken/Hebei/1/96(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536724 Influenza A virus
(A/Chicken/Hebei/2/98(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536725 Influenza A virus
(A/Chicken/Hebei/3/98(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536726 Influenza A virus
(A/Chicken/Henan/98(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536727 Influenza A virus
(A/Chicken/Liaoning/99(H9N2)) nonfunctional matrix protein gene,
partial sequence. AF536728 Influenza A virus
(A/Chicken/Shandong/98(H9N2)) nonfunctional matrix protein gene,
partial sequence. AJ291398 Influenza A virus
(A/Chicken/Pakistan/2/99 (H9N2)) M1 gene for Matrix Protein 1 (exon
1) and M2 gene for Matrix Protein 2 (exons 1 and 2), genomic RNA
AJ427865 Influenza A virus (A/quail/Hong Kong/FY298/00 (H9N2))
partial m gene for matrix protein, genomic RNA AY180461 Influenza A
virus strain A/Pigeon/Nanchang/2-0461/2000 (H9N2) matrix protein
(M) gene, partial cds. AY180462 Influenza A virus strain
A/Duck/Nanchang/11-290/2000 (H9N2) matrix protein (M) gene, partial
cds. AY180463 Influenza A virus strain A/Duck/Nanchang/11-197/2000
(H9N2) matrix protein (M) gene, partial cds. AY180464 Influenza A
virus strain A/Duck/Nanchang/11-392/2000 (H9N2) matrix protein (M)
gene, partial cds. AY180477 Influenza A virus strain
A/Chicken/Nanchang/4-361/2001 (H9N2) matrix protein (M) gene,
partial cds. AY180485 Influenza A virus strain
A/Pigeon/Nanchang/11-145/2000 (H9N2) matrix protein (M) gene,
partial cds. AY180486 Influenza A virus strain
A/Duck/Nanchang/1-0070/2000 (H9N2) matrix protein (M) gene, partial
cds. AY180489 Influenza A virus strain A/Duck/Nanchang/10-389/2000
(H9N2) matrix protein (M) gene, partial cds. AY180490 Influenza A
virus strain A/Chicken/Nanchang/1-0016/2000 (H9N2) matrix protein
(M) gene, partial cds. AY180492 Influenza A virus strain
A/Pigeon/Nanchang/7-058/2000 (H9N2) matrix protein (M) gene,
partial cds. AY180495 Influenza A virus strain
A/Quail/Nanchang/2-0460/2000 (H9N2) matrix protein (M) gene,
partial cds. AY180502 Influenza A virus strain
A/Chicken/Nanchang/4-010/2000 (H9N2) matrix protein (M) gene,
partial cds. AY180504 Influenza A virus strain
A/Quail/Nanchang/4-040/2000 (H9N2) matrix protein (M) gene, partial
cds. AY180506 Influenza A virus strain
A/Chicken/Nanchang/4-301/2001 (H9N2) matrix protein (M) gene,
partial cds. AY180516 Influenza A virus strain
A/Duck/Nanchang/7-092/2000 (H9N2) matrix protein (M) gene, partial
cds. AY180519 Influenza A virus strain A/Wild
Duck/Nanchang/2-0480/2000 (H9N2) matrix protein (M) gene, partial
cds. AY253755 Influenza A virus (A/Chicken/Shanghai/F/98(H9N2))
matrix protein M1 and membrane ion channel M2 genes, complete cds.
AY496852 Influenza A virus (A/chicken/Mudanjiang/0823/2000(H9N2))
matrix protein (M1) mRNA, complete cds. AY633165 Influenza A virus
(A/mallard/Alberta/17/91(H9N2)) matrix protein (M) gene, complete
cds. AY633277 Influenza A virus (A/mallard/Alberta/321/88(H9N2))
matrix protein (M) gene, complete cds. AY633293 Influenza A virus
(A/mallard/Alberta/11/91(H9N2)) matrix protein (M) gene, complete
cds. AY664464 Influenza A virus (A/shorebird/Delaware/276/99
(H9N2)) nonfunctional matrix protein mRNA, partial sequence.
AY664679 Influenza A virus (A/chicken/HongKong/CSW153/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664680 Influenza A virus
(A/chicken/HongKong/AP45/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664681 Influenza A virus (A/chicken/HongKong/BD90/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664682 Influenza A virus
(A/chicken/HongKong/CSW291/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664683 Influenza A virus (A/chicken/HongKong/CSW304/03(H9N2))
membrane protein M2 (M) and membrane protein M1 (M) genes, partial
cds. AY664684 Influenza A virus (A/chicken/HongKong/FY23/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664685 Influenza A virus
(A/guineafowl/HongKong/NT101/03(H9N2)) membrane protein M2 (M)
gene, partial cds; and membrane protein M1 (M) gene, complete cds.
AY664686 Influenza A virus (A/chicken/HongKong/NT142/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664687 Influenza A virus
(A/chicken/HongKong/SF1/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664688 Influenza A virus (A/chicken/HongKong/SSP101/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664689 Influenza A virus
(A/chicken/HongKong/TP38/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664690 Influenza A virus (A/chicken/HongKong/WF126/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664691 Influenza A virus
(A/pigeon/HongKong/WF53/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664692 Influenza A virus (A/pheasant/HongKong/WF54/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664693 Influenza A virus
(A/guineafowl/HongKong/NT184/03(H9N2)) membrane protein M2 (M)
gene, partial cds; and membrane protein M1 (M) gene, complete cds.
AY664694 Influenza A virus (A/chicken/HongKong/WF120/03(H9N2))
membrane protein M2 (M) and membrane protein M1 (M) genes, partial
cds. AY664695 Influenza A virus (A/chicken/HongKong/NT366/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY664696 Influenza A virus
(A/chicken/HongKong/SSP418/03(H9N2)) membrane protein M2 (M) gene,
partial cds; and membrane protein M1 (M) gene, complete cds.
AY664697 Influenza A virus (A/chicken/HongKong/YU427/03(H9N2))
membrane protein M2 (M) gene, partial cds; and membrane protein M1
(M) gene, complete cds. AY800234 Influenza A virus
(A/chicken/Korea/S1/2003(H9N2)) matrix protein (M) gene, complete
cds. AY862614 Influenza A virus (A/silky chicken/Korea/S3/03(H9N2))
matrix protein (M) gene, complete cds. AY862615 Influenza A virus
(A/chicken/Korea/S4/03(H9N2)) matrix protein (M) gene, complete
cds. AY862616 Influenza A virus (A/chicken/Korea/S5/03(H9N2))
matrix protein (M) gene, complete cds. AY862617 Influenza A virus
(A/chicken/Korea/S12/03(H9N2)) matrix protein (M) gene, complete
cds. AY862618 Influenza A virus (A/duck/Korea/S13/03(H9N2)) matrix
protein (M) gene, complete cds. AY862619 Influenza A virus
(A/dove/Korea/S14/03(H9N2)) matrix protein (M) gene, partial cds.
AY862620 Influenza A virus (A/chicken/Korea/S15/03(H9N2)) matrix
protein (M) gene, complete cds. AY862621 Influenza A virus
(A/chicken/Korea/S16/03(H9N2)) matrix protein (M) gene, complete
cds. AY862622 Influenza A virus (A/chicken/Korea/S18/03(H9N2))
matrix protein (M) gene, complete cds. AF156458 Influenza A virus
(A/Chicken/Hong Kong/G9/97(H9N2)) segment 7 matrix protein M1 (M1)
and matrix protein M2 (M2) genes, complete cds. AF156459 Influenza
A virus (A/Chicken/Hong Kong/G23/97(H9N2)) segment 7 matrix protein
M1 (M1) and matrix protein M2 (M2) genes, complete cds. AF156460
Influenza A virus (A/Pigeon/Hong Kong/Y233/97(H9N2)) segment 7
matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete
cds. AF156461 Influenza A virus (A/Duck/Hong Kong/Y280/97(H9N2))
segment 7 matrix protein M1 (M1) and matrix protein M2 (M2) genes,
complete cds. AF156462 Influenza A virus (A/Duck/Hong
Kong/Y439/97(H9N2)) segment 7
matrix protein M1 (M1) and matrix protein M2 (M2) genes, complete
cds. AF156463 Influenza A virus (A/Quail/Hong Kong/G1/97 (H9N2))
segment 7 matrix protein M1 (M1) gene, complete cds; and matrix
protein M2 (M2) gene, partial cds. AF156464 Influenza A virus
(A/Chicken/Hong Kong/739/94(H9N2)) segment 7 matrix protein M1 (M1)
gene, complete cds; and matrix protein M2 (M2) gene, partial cds.
AF156465 Influenza A virus (A/Quail/Hong Kong/AF157/92(H9N2))
segment 7 matrix protein M1 (M1) gene, complete cds; and matrix
protein M2 (M2) gene, partial cds. AF156466 Influenza A virus
(A/Chicken/Beijing/1/94(H9N2)) segment 7 matrix protein M1 (M1)
gene, complete cds; and matrix protein M2 (M2) gene, partial cds.
AF156467 Influenza A virus (A/Chicken/Korea/38349-p96323/96(H9N2))
segment 7 matrix protein M1 (M1) gene, complete cds; and matrix
protein M2 (M2) gene, partial cds. AF156468 Influenza A virus
(A/Chicken/Korea/25232-96006/96(H9N2)) segment 7 matrix protein M1
(M1) gene, complete cds; and matrix protein M2 (M2) gene, partial
cds. AF156469 Influenza A virus (A/Shorebird/Delaware/9/96(H9N2))
segment 7 matrix protein M1 (M1) gene, complete cds; and matrix
protein M2 (M2) gene, partial cds. AF156470 Influenza A virus
(A/Quail/Arkansas/29209-1/93(H9N2)) segment 7 matrix protein M1
(M1) gene, complete cds; and matrix protein M2 (M2) gene, partial
cds. AF156471 Influenza A virus (A/Turkey/California/189/66(H9N2))
segment 7 matrix protein M1 (M1) gene, complete cds; and matrix
protein M2 (M2) gene, partial cds. AF203788 Influenza A virus
(A/Chicken/Korea/MS96/96(H9N2)) matrix protein 1 mRNA, complete
cds; and matrix protein 2 mRNA, partial cds. AF222662 Influenza A
virus (A/Quail/Hong Kong/A17/99(H9N2)) segment 7 M1 (M1) gene,
partial cds. AF222663 Influenza A virus (A/Pigeon/Hong
Kong/FY6/99(H9N2)) segment 7 M1 (M1) gene, partial cds. AF222664
Influenza A virus (A/Chicken/Hong Kong/NT16/99(H9N2)) segment 7 M1
(M1) gene, partial cds. AF222665 Influenza A virus (A/Quail/Hong
Kong/SSP10/99(H9N2)) segment 7 M1 (M1) gene, partial cds. AF222666
Influenza A virus (A/Pheasant/Hong Kong/SSP11/99(H9N2)) segment 7
M1 (M1) gene, partial cds. AF222667 Influenza A virus
(A/Chicken/Hong Kong/FY20/99(H9N2)) segment 7 M1 (M1) gene, partial
cds. AF222668 Influenza A virus (A/Chicken/Hong Kong/KC12/99(H9N2))
segment 7 M1 (M1) gene, partial cds. AF222669 Influenza A virus
(A/Quail/Hong Kong/NT28/99(H9N2)) segment 7 M1 (M1) gene, partial
cds. AF222670 Influenza A virus (A/Chicken/Hong Kong/SF2/99(H9N2))
segment 7 M1 (M1) gene, partial cds. Sequences used in analysis of
Influenza A Nucleocapsid Protein (NP) AF156415 Influenza A virus
(A/Turkey/California/189/66(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF523423 Influenza A virus (A/Duck/Hong
Kong/86/76(H9N2)) nucleocapsid protein (NP) gene, complete cds.
AF523424 Influenza A virus (A/Duck/Hong Kong/366/78(H9N2))
nucleocapsid protein (NP) gene, partial cds. AF523421 Influenza A
virus (A/Duck/Hong Kong/289/78(H9N2)) nucleocapsid protein (NP)
gene, complete cds. AF523422 Influenza A virus (A/Duck/Hong
Kong/552/79(H9N2)) nucleocapsid protein (NP) gene, partial cds.
AY633279 Influenza A virus (A/mallard/Alberta/321/88(H9N2))
nucleoprotein (NP) gene, complete cds. AY633295 Influenza A virus
(A/mallard/Alberta/11/91(H9N2)) nucleoprotein (NP) gene, partial
cds. AY633167 Influenza A virus (A/mallard/Alberta/17/91(H9N2))
nucleoprotein (NP) gene, complete cds. AF156410 Influenza A virus
(A/Quail/Hong Kong/AF157/92(H9N2)) segment 5 nucleoprotein (NP)
gene, complete cds. AF156414 Influenza A virus
(A/Quail/Arkansas/29209-1/93 (H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF156408 Influenza A virus (A/Chicken/Hong
Kong/739/94(H9N2)) segment 5 nucleoprotein (NP) gene, complete cds.
AF156409 Influenza A virus (A/Chicken/Beijing/1/94(H9N2)) segment 5
nucleoprotein (NP) gene, complete cds. AF536699 Influenza A virus
(A/Chicken/Beijing/1/95(H9N2)) nucleoprotein (NP) gene, partial
cds. AF508596 Influenza A virus (A/Ostrich/South
Africa/9508103/95(H9N2)) segment 5 nucleoprotein (NP) gene, partial
cds. AF508600 Influenza A virus (A/Duck/Germany/113/95(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AB020778 Influenza
A virus gene for nucleoprotein, complete cds. AF508613 Influenza A
virus (A/Chicken/Shandong/6/96(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508617 Influenza A virus
(A/Quail/Shanghai/8/96(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF536703 Influenza A virus
(A/Chicken/Hebei/1/96(H9N2)) nucleoprotein (NP) gene, partial cds.
AF156411 Influenza A virus (A/Chicken/Korea/38349-96323/96 (H9N2))
segment 5 nucleoprotein (NP) gene, complete cds. AF156412 Influenza
A virus (A/Chicken/Korea/25232-96006/96 (H9N2)) segment 5
nucleoprotein (NP) gene, partial cds. M63779 Influenza
A/FPV/Dobson/`Dutch`/27 (H7N7) nucleoprotein mRNA, complete cds.
M63784 Influenza A/Teal/Iceland/29/80 (H7N7) nucleoprotein mRNA,
complete cds. AJ620352 Influenza A virus
(A/Chicken/Germany/R28/03(H7N7)) NP gene for nucleoprotein, genomic
RNA. AY342425 Influenza A virus (A/Netherlands/219/03(H7N7))
nucleocapsid protein gene, complete cds. AY342426 Influenza A virus
(A/Netherlands/033/03(H7N7)) nucleocapsid protein gene, complete
cds. AY342427 Influenza A virus (A/chicken/Netherlands/1/03(H7N7))
nucleocapsid protein gene, complete cds. AF156413 Influenza A virus
(A/Shorebird/Delaware/9/96 (H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF203787 Influenza A virus
(A/Chicken/Korea/MS96/96(H9N2)) nucleoprotein mRNA, complete cds.
AF156402 Influenza A virus (A/Chicken/Hong Kong/G9/97(H9N2))
segment 5 nucleoprotein (NP) gene, complete cds. AF156403 Influenza
A virus (A/Chicken/Hong Kong/G23/97(H9N2)) segment 5 nucleoprotein
(NP) gene, complete cds. AF156404 Influenza A virus (A/Pigeon/Hong
Kong/Y233/97(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds.
AF156405 Influenza A virus (A/Duck/Hong Kong/Y280/97(H9N2)) segment
5 nucleoprotein (NP) gene, partial cds. AF156406 Influenza A virus
(A/Duck/Hong Kong/Y439/97(H9N2)) segment 5 nucleoprotein (NP) gene,
complete cds. AF156407 Influenza A virus (A/Quail/Hong Kong/G1/97
(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds. AF508612
Influenza A virus (A/Chicken/Sichuan/5/97(H9N2)) segment 5
nucleoprotein (NP) gene, partial cds. AF536702 Influenza A virus
(A/Chicken/Guangdong/97(H9N2)) nucleoprotein (NP) gene, partial
cds. AF536700 Influenza A virus (A/Chicken/Beijing/2/97(H9N2))
nucleoprotein (NP) gene, partial cds. AF508615 Influenza A virus
(A/Chicken/Shenzhen/9/97(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF508616 Influenza A virus (A/Duck/Nanjing/1/97(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AB049161 Influenza
A virus (A/parakeet/Chiba/1/97(H9N2)) NP gene for nucleoprotein,
complete cds. AF508603 Influenza A virus
(A/Pheasant/Ireland/PV18/97(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508607 Influenza A virus
(A/Chicken/Guangdong/11/97(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508609 Influenza A virus
(A/Chicken/Heilongjiang/10/97(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508608 Influenza A virus
(A/Chicken/Hebei/4/98(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF508605 Influenza A virus
(A/Chicken/Beijing/8/98(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF508599 Influenza A virus (A/Chicken/Germany/R45/98
(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds. AF536708
Influenza A virus (A/Chicken/Shandong/98(H9N2)) nucleoprotein (NP)
gene, partial cds. AY253753 Influenza A virus
(A/Chicken/Shanghai/F/98(H9N2)) nucleoprotein (NP) gene, complete
cds. AF536704 Influenza A virus (A/Chicken/Hebei/2/98(H9N2))
nucleoprotein (NP) gene, partial cds. AF536705 Influenza A virus
(A/Chicken/Hebei/3/98(H9N2)) nucleoprotein (NP) gene, partial cds.
AF536706 Influenza A virus (A/Chicken/Henan/98(H9N2)) nucleoprotein
(NP) gene, partial cds. AB049162 Influenza A virus
(A/parakeet/Narita/92A/98(H9N2)) NP gene for nucleoprotein,
complete cds. AF186270 Influenza A virus (A/Quail/Hong
Kong/NT28/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds.
AF186271 Influenza A virus (A/Silkie Chicken/Hong
Kong/SF43/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds.
AF186272 Influenza A virus (A/Chicken/Hong Kong/SF2/99(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AF222614 Influenza
A virus (A/Quail/Hong Kong/A17/99(H9N2)) segment 5 nucleoprotein
(NP) gene, partial cds. AF222615 Influenza A virus (A/Pigeon/Hong
Kong/FY6/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds.
AF222616 Influenza A virus (A/Chicken/Hong Kong/NT16/99(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AF222617 Influenza
A virus (A/Quail/Hong Kong/SSP10/99(H9N2)) segment 5 nucleoprotein
(NP) gene, partial cds. AF222618 Influenza A virus (A/Pheasant/Hong
Kong/SSP11/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial
cds. AF536707 Influenza A virus (A/Chicken/Liaoning/99(H9N2))
nucleoprotein (NP) gene, partial cds. AJ291394 Influenza A virus
(A/Chicken/Pakistan/2/99 (H9N2)) NP gene for Nucleoprotein, genomic
RNA. AF536701 Influenza A virus (A/Chicken/Beijing/3/99(H9N2))
nucleoprotein (NP) gene, partial cds. AF508611 Influenza A virus
(A/Chicken/Ningxia/5/99(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF508614 Influenza A virus
(A/Chicken/Shijiazhuang/2/99(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508604 Influenza A virus
(A/Chicken/Korea/99029/99(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF222619 Influenza A virus (A/Chicken/Hong
Kong/FY20/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial cds.
AF222620 Influenza A virus (A/Chicken/Hong Kong/KC12/99(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AF222621 Influenza
A virus (A/Silky Chicken/Hong Kong/SF44/99(H9N2)) segment 5
nucleoprotein (NP) gene, partial cds. AF508601 Influenza A virus
(A/Chicken/Iran/11T/99(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF508602 Influenza A virus (A/Chicken/Saudi
Arabia/532/99(H9N2)) segment 5 nucleoprotein (NP) gene, partial
cds. AF508597 Influenza A virus (A/Chicken/Pakistan/4/99(H9N2))
segment 5 nucleoprotein (NP) gene, partial cds. AF508598 Influenza
A virus (A/Chicken/Pakistan/5/99(H9N2)) segment 5 nucleoprotein
(NP) gene, partial cds. AF508606 Influenza A virus
(A/Chicken/Guangdong/10/00(H9N2)) segment 5 nucleoprotein (NP)
gene, partial cds. AF508610 Influenza A virus
(A/Chicken/Henan/62/00(H9N2)) segment 5 nucleoprotein (NP) gene,
partial cds. AF523410 Influenza A virus
(A/Duck/Shantou/1043/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523411 Influenza A virus
(A/Duck/Shantou/2134/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523413 Influenza A virus
(A/Duck/Shantou/1042/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523415 Influenza A virus
(A/Duck/Shantou/2102/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523416 Influenza A virus
(A/Duck/Shantou/830/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523417 Influenza A virus
(A/Duck/Shantou/2144/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523419 Influenza A virus
(A/Duck/Shantou/2143/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523420 Influenza A virus
(A/Duck/Shantou/1881/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AJ427864 Influenza A virus (A/quail/Hong Kong/FY298/00
(H9N2)) partial np gene for nucleoprotein, genomic RNA AY180525
Influenza A virus (A/Pigeon/Nanchang/2-0461/2000(H9N2))
nucleoprotein (NP) gene, partial cds. AY180534 Influenza A virus
strain A/Duck/Nanchang/7-092/2000 (H9N2) nucleoprotein (NP) gene,
partial cds. AY180537 Influenza A virus strain
A/Duck/Nanchang/11-392/2000 (H9N2) nucleoprotein (NP) gene, partial
cds. AY180538 Influenza A virus
(A/Pigeon/Nanchang/11-145/2000(H9N2)) nucleoprotein (NP) gene,
partial cds. AY180542 Influenza A virus strain
A/Duck/Nanchang/11-197/2000 (H9N2) nucleoprotein (NP) gene, partial
cds. AY180544 Influenza A virus strain A/Duck/Nanchang/11-290/2000
(H9N2) nucleoprotein (NP) gene, partial cds. AY180560 Influenza A
virus (A/Pigeon/Nanchang/7-058/2000(H9N2)) nucleoprotein (NP) gene,
partial cds. AY180562 Influenza A virus strain
A/Chicken/Nanchang/4-010/2000 (H9N2) nucleoprotein (NP) gene,
partial cds. AY180563 Influenza A virus strain
A/Quail/Nanchang/4-040/2000 (H9N2) nucleoprotein (NP) gene, partial
cds. AY180564 Influenza A virus (A/Wild
Duck/Nanchang/2-0480/2000(H9N2)) nucleoprotein (NP) gene, partial
cds. AY180575 Influenza A virus
(A/Quail/Nanchang/2-0460/2000(H9N2)) nucleoprotein (NP) gene,
partial cds. AY496851 Influenza A virus
(A/chicken/Mudanjiang/0823/2000(H9N2)) nucleoprotein (np) mRNA,
complete cds. AY180581 Influenza A virus
(A/Chicken/Nanchang/1-0016/2000(H9N2))
nucleoprotein (NP) gene, partial cds. AY180583 Influenza A virus
strain A/Duck/Nanchang/10-389/2000 (H9N2) nucleoprotein (NP) gene,
partial cds. AY180584 Influenza A virus strain
A/Duck/Nanchang/1-0070/2000 (H9N2) nucleoprotein (NP) gene, partial
cds. AY768567 Influenza A virus (A/chicken/Korea/SNU0028/00(H9N2))
nucleocapsid protein (NP) gene, partial cds. AY768568 Influenza A
virus (A/chicken/Korea/SNU0037/00(H9N2)) nucleocapsid protein (NP)
gene, partial cds. AY768569 Influenza A virus
(A/chicken/Korea/SNU0057/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY768570 Influenza A virus
(A/chicken/Korea/SNU0073/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY768571 Influenza A virus
(A/chicken/Korea/SNU0091/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY768572 Influenza A virus
(A/chicken/Korea/SNU0140/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY768573 Influenza A virus
(A/chicken/Korea/SNU0146/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY768574 Influenza A virus
(A/chicken/Korea/SNU1035C/00(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY268949 Influenza A virus
(A/chicken/Wangcheng/4/2001(H9N2)) nucleoprotein mRNA, complete
cds. AY180578 Influenza A virus strain
A/Chicken/Nanchang/4-301/2001 (H9N2) nucleoprotein (NP) gene,
partial cds. AY180551 Influenza A virus
(A/Chicken/Nanchang/4-361/2001(H9N2)) nucleoprotein (NP) gene,
partial cds. AF523418 Influenza A virus
(A/Duck/Shantou/2088/01(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523414 Influenza A virus
(A/Duck/Shantou/1605/01(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AF523412 Influenza A virus (A/Wild
Duck/Shantou/4808/01(H9N2)) nucleocapsid protein (NP) gene, partial
cds. AY800236 Influenza A virus (A/chicken/Korea/S1/2003(H9N2))
nucleoprotein (NP) gene, partial cds. AY862646 Influenza A virus
(A/silky chicken/Korea/S3/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862647 Influenza A virus
(A/chicken/Korea/S4/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862648 Influenza A virus
(A/chicken/Korea/S5/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862649 Influenza A virus
(A/chicken/Korea/S12/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862650 Influenza A virus (A/Duck/Korea/S13/03(H9N2))
nucleocapsid protein (NP) gene, partial cds. AY862651 Influenza A
virus (A/dove/Korea/S14/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862652 Influenza A virus
(A/chicken/Korea/S15/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862653 Influenza A virus
(A/chicken/Korea/S16/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY862654 Influenza A virus
(A/chicken/Korea/S18/03(H9N2)) nucleocapsid protein (NP) gene,
partial cds. AY664717 Influenza A virus
(A/chicken/HongKong/CSW153/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664718 Influenza A virus
(A/chicken/HongKong/AP45/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664719 Influenza A virus
(A/chicken/HongKong/BD90/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664720 Influenza A virus
(A/chicken/HongKong/CSW291/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664721 Influenza A virus
(A/chicken/HongKong/CSW304/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664722 Influenza A virus
(A/chicken/HongKong/FY23/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664723 Influenza A virus
(A/guineafowl/HongKong/NT101/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664724 Influenza A virus
(A/chicken/HongKong/NT142/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664725 Influenza A virus
(A/chicken/HongKong/SF1/03(H9N2)) nucleoprotein (NP) gene, complete
cds. AY664726 Influenza A virus
(A/chicken/HongKong/SSP101/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664727 Influenza A virus
(A/chicken/HongKong/TP38/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664728 Influenza A virus
(A/chicken/HongKong/WF126/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664729 Influenza A virus
(A/pigeon/HongKong/WF53/03(H9N2)) nucleoprotein (NP) gene, complete
cds. AY664730 Influenza A virus (A/pheasant/HongKong/WF54/03(H9N2))
nucleoprotein (NP) gene, complete cds. AY664731 Influenza A virus
(A/guineafowl/HongKong/NT184/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664732 Influenza A virus
(A/chicken/HongKong/WF120/03(H9N2)) nucleoprotein (NP) gene,
complete cds. AY664733 Influenza A virus
(A/chicken/HongKong/NT366/03(H9N2)) nucleoprotein (NP) gene,
partial cds. AY664734 Influenza A virus
(A/chicken/HongKong/SSP418/03(H9N2)) nucleoprotein (NP) gene,
partial cds. AY664735 Influenza A virus
(A/chicken/HongKong/YU427/03(H9N2)) nucleoprotein (NP) gene,
partial cds. AY788915 Influenza A virus
(A/chicken/China/HSS2004(H9N2)) nucleoprotein (NP) gene, complete
cds. AY586423 Influenza A virus (A/mallard/Italy/33/01(H7N3))
nucleoprotein gene, partial cds. AY586424 Influenza A Virus
(A/mallard/Italy/43/01(H7N3)) nucleoprotein gene, partial cds.
AY586425 Influenza A virus (A/turkey/Italy/220158/2002(H7N3))
nucleoprotein gene, partial cds. AY586426 Influenza A Virus
(A/turkey/Italy/214845/02(H7N3)) nucleoprotein gene, partial cds.
AJ627486 Influenza A virus (A/turkey/Italy/214845/2002(H7N3)) NP
gene for nucleoprotein, genomic RNA. AJ627495 Influenza A virus
(A/turkey/Italy/220158/2002(H7N3)) NP gene for nucleoprotein,
genomic RNA. AY303658 Influenza A virus
(A/chicken/Chile/176822/02(H7N3)) nucleoprotein gene, complete cds.
AY303659 Influenza A virus (A/chicken/Chile/4957/02(H7N3))
nucleoprotein gene, complete cds. AY611527 Influenza A virus
(A/chicken/British Columbia/04(H7N3)) nucleoprotein (NP) gene,
complete cds. AY646081 Influenza A virus (A/chicken/British
Columbia/GSC_human_B/04(H7N3)) nucleoprotein (NP) gene, complete
cds. AY648290 Influenza A virus (A/GSC_chicken_B/British
Columbia/04(H7N3)) nucleoprotein (NP) gene, complete cds. AY650273
Influenza A virus (A/GSC_chicken/British Columbia/04(H7N3))
nucleoprotein (NP) gene, complete cds. AF144303 Influenza A virus
(A/Goose/Guangdong/1/96(H5N1)) nucleocapsid protein (NP) gene,
complete cds. AF046084 Influenza A virus (A/Chicken/Hong
Kong/220/97 (H5N1)) nucleoprotein gene, complete cds. AF057293
Influenza A virus (A/chicken/Hong Kong/258/97(H5N1)) nucleoprotein
mRNA, complete cds. AF098617 Influenza A virus (A/Chicken/Hong
Kong/y388/97 (H5N1)) nucleoprotein (NP) gene, complete cds.
AF098618 Influenza A virus (A/Chicken/Hong Kong/728/97 (H5N1))
nucleoprotein (NP) gene, complete cds. AF098619 Influenza A virus
(A/Chicken/Hong Kong/786/97 (H5N1)) nucleoprotein (NP) gene,
complete cds. AF098620 Influenza A virus (A/Chicken/Hong
Kong/915/97 (H5N1)) nucleoprotein (NP) gene, complete cds. AF098621
Influenza A virus (A/Duck/Hong Kong/p46/97 (H5N1)) nucleoprotein
(NP) gene, complete cds. AF098622 Influenza A virus (A/Duck/Hong
Kong/y283/97 (H5N1)) nucleoprotein (NP) gene, complete cds.
AF098623 Influenza A virus (A/Goose/Hong Kong/w355/97 (H5N1))
nucleoprotein (NP) gene, complete cds. AF370122 Influenza A virus
(A/Goose/Guangdong/3/97(H5N1)) nucleoprotein gene, complete cds.
AF216712 Influenza A virus (A/Environment/Hong Kong/437-4/99
(H5N1)) nucleoprotein gene, complete cds. AF216720 Influenza A
virus (A/Environment/Hong Kong/437-6/99 (H5N1)) nucleoprotein gene,
complete cds. AF216728 Influenza A virus (A/Environment/Hong
Kong/437-8/99 (H5N1)) nucleoprotein gene, complete cds. AF216736
Influenza A virus (A/Environment/Hong Kong/437-10/99 (H5N1))
nucleoprotein gene, complete cds. AY585429 Influenza A virus
(A/duck/Guangxi/07/1999(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585439 Influenza A virus (A/duck/Zhejiang/11/2000(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585440 Influenza A virus
(A/duck/Zhejiang/52/2000(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585428 Influenza A virus (A/duck/Guangdong/40/2000(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585423 Influenza A virus
(A/duck/Fujian/19/2000(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585425 Influenza A virus (A/duck/Guangdong/07/2000(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585426 Influenza A virus
(A/duck/Guangdong/12/2000(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY059492 Influenza A virus (A/Goose/Hong Kong/ww26/2000(H5N1))
segment 5 nucleocapsid protein (NP) gene, partial cds. AY059493
Influenza A virus (A/Goose/Hong Kong/ww28/2000(H5N1)) segment 5
nucleocapsid protein (NP) gene, partial cds. AY059494 Influenza A
virus (A/Duck/Hong Kong/ww381/2000(H5N1)) segment 5 nucleocapsid
protein (NP) gene, partial cds. AY059495 Influenza A virus
(A/Duck/Hong Kong/ww461/2000(H5N1)) segment 5 nucleocapsid protein
(NP) gene, partial cds. AY059496 Influenza A virus (A/Goose/Hong
Kong/ww491/2000(H5N1)) segment 5 nucleocapsid protein (NP) gene,
partial cds. AY059497 Influenza A virus (A/Duck/Hong
Kong/2986.1/2000(H5N1)) segment 5 nucleocapsid protein (NP) gene,
partial cds. AY059498 Influenza A virus (A/Goose/Hong
Kong/3014.8/2000(H5N1)) segment 5 nucleocapsid protein (NP) gene,
partial cds. AF398419 Influenza A virus (A/Goose/Hong
Kong/385.3/2000(H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF398420 Influenza A virus (A/Goose/Hong Kong/385.5/2000(H5N1))
nucleocapsid protein (NP) gene, partial cds. AF468842 Influenza A
virus (A/Duck/Anyang/AVL-1/2001(H5N1)) nucleoprotein (NP) gene,
complete cds. AF509117 Influenza A virus (A/Chicken/Hong
Kong/FY77/01 (H5N1)) nucleocapsid protein (NP) gene, complete cds.
AF509118 Influenza A virus (A/Chicken/Hong Kong/YU562/01 (H5N1))
nucleocapsid protein (NP) gene, complete cds. AF509119 Influenza A
virus (A/Chicken/Hong Kong/YU563/01 (H5N1)) nucleocapsid protein
(NP) gene, complete cds. AY585438 Influenza A virus
(A/duck/Shanghai/38/2001(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY221548 Influenza A virus
(A/Chicken/HongKong/NT873.3/01-MB(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221549 Influenza A virus
(A/Chicken/HongKong/NT873.3/01(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221550 Influenza A virus
(A/Chicken/HongKong/FY150/01-MB(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221551 Influenza A virus
(A/Chicken/HongKong/FY150/01(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY221552 Influenza A virus
(A/Pheasant/HongKong/FY155/01-MB(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221553 Influenza A virus
(A/Pheasant/HongKong/FY155/01(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221554 Influenza A virus
(A/Chicken/HongKong/YU822.2/01-MB(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221555 Influenza A virus
(A/Chicken/HongKong/YU822.2/01(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY221556 Influenza A virus
(A/Chicken/HongKong/YU562/01(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AF509120 Influenza A virus (A/Chicken/Hong
Kong/FY150/01 (H5N1)) nucleocapsid protein (NP) gene, complete cds.
AF509121 Influenza A virus (A/Pheasant/Hong Kong/FY155/01 (H5N1))
nucleocapsid protein (NP) gene, complete cds. AF509122 Influenza A
virus (A/Silky Chicken/Hong Kong/SF189/01 (H5N1)) nucleocapsid
protein (NP) gene, partial cds. AF509123 Influenza A virus
(A/Quail/Hong Kong/SF203/01 (H5N1)) nucleocapsid protein (NP) gene,
partial cds. AF509124 Influenza A virus (A/Pigeon/Hong
Kong/SF215/01 (H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF509125 Influenza A virus (A/Chicken/Hong Kong/SF219/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509126 Influenza A
virus (A/Chicken/Hong Kong/715.5/01 (H5N1)) nucleocapsid protein
(NP) gene, partial cds. AF509127 Influenza A virus (A/Chicken/Hong
Kong/751.1/01 (H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF509128 Influenza A virus (A/Chicken/Hong Kong/822.1/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509129 Influenza A
virus (A/Chicken/Hong Kong/829.2/01 (H5N1)) nucleocapsid protein
(NP) gene, partial cds. AF509130 Influenza A virus (A/Chicken/Hong
Kong/830.2/01 (H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF509131 Influenza A virus (A/Chicken/Hong Kong/858.3/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509132 Influenza A
virus (A/Chicken/Hong Kong/866.3/01 (H5N1)) nucleocapsid protein
(NP) gene, partial cds. AF509133 Influenza A virus (A/Chicken/Hong
Kong/867.1/01 (H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF509134 Influenza A virus (A/Chicken/Hong Kong/879.1/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509135 Influenza A
virus (A/Chicken/Hong Kong/873.3/01 (H5N1)) nucleocapsid protein
(NP) gene, partial cds. AF509136 Influenza A virus (A/Chicken/Hong
Kong/876.1/01 (H5N1)) nucleocapsid protein (NP) gene, partial cds.
AF509137 Influenza A virus (A/Chicken/Hong Kong/891.1/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509138 Influenza A
virus (A/Chicken/Hong Kong/893.2/01 (H5N1)) nucleocapsid protein
(NP) gene, partial cds. AF509139 Influenza A virus (A/Goose/Hong
Kong/76.1/01 (H5N1)) nucleocapsid protein (NP) gene, complete cds.
AF509140 Influenza A virus (A/Goose/Hong Kong/ww100/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AF509141 Influenza A
virus (A/Duck/Hong Kong/573.4/01 (H5N1)) nucleocapsid protein (NP)
gene, partial cds.
AF509142 Influenza A virus (A/Duck/Hong Kong/646.3/01 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AY585424 Influenza A
virus (A/duck/Guangdong/01/2001(H5N1)) nucleoprotein (NP) mRNA,
complete cds. AY585422 Influenza A virus
(A/duck/Fujian/17/2001(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585430 Influenza A virus (A/duck/Guangxi/22/2001(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585431 Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585432 Influenza A virus (A/duck/Guangxi/50/2001(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585434 Influenza A virus
(A/duck/Shanghai/08/2001(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585435 Influenza A virus (A/duck/Shanghai/13/2001(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585436 Influenza A virus
(A/duck/Shanghai/35/2002(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585437 Influenza A virus (A/duck/Shanghai/37/2002(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585433 Influenza A virus
(A/duck/Guangxi/53/2002(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585427 Influenza A virus (A/duck/Guangdong/22/2002(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY585420 Influenza A virus
(A/duck/Fujian/01/2002(H5N1)) nucleoprotein (NP) mRNA, complete
cds. AY585421 Influenza A virus (A/duck/Fujian/13/2002(H5N1))
nucleoprotein (NP) mRNA, complete cds. AY575907 Influenza A virus
(A/Gs/HK/739.2/02 (H5N1)) nucleocapsid protein (NP) gene, partial
cds. AY575908 Influenza A virus (A/Eg/HK/757.3/02 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AY575909 Influenza A
virus (A/G.H/HK/793.1/02 (H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY575910 Influenza A virus (A/Dk/HK/821/02 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AY575911 Influenza A
virus (A/Ck/HK/31.4/02 (H5N1)) nucleocapsid protein (NP) gene,
complete cds. AY575912 Influenza A virus (A/Ck/HK/61.9/02 (H5N1))
nucleocapsid protein (NP) gene, complete cds. AY575913 Influenza A
virus (A/Ck/HK/YU777/02 (H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY575914 Influenza A virus (A/Ck/HK/96.1/02 (H5N1))
nucleocapsid protein (NP) gene, partial cds. AY575915 Influenza A
virus (A/Ck/HK/409.1/02 (H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY575916 Influenza A virus (A/Ph/HK/sv674.15/02
(H5N1)) nucleocapsid protein (NP) gene, partial cds. DQ023146
Influenza A virus (A/chicken/sd/1/02(H5N1)) nucleoprotein (NP)
mRNA, complete cds. AY676037 Influenza A virus (A/duck/Hong
Kong/821/02(H5N1)) nucleoprotein (NP) gene, complete cds. AY651510
Influenza A virus (A/Gf/HK/38/2002(H5N1)) nucleocapsid protein (NP)
gene, complete cds. AY651511 Influenza A virus
(A/Ck/HK/31.2/2002(H5N1)) nucleocapsid protein (NP) gene, complete
cds. AY651512 Influenza A virus (A/Ck/HK/37.4/2002(H5N1))
nucleocapsid protein (NP) gene, complete cds. AY651513 Influenza A
virus (A/SCk/HK/YU100/2002(H5N1)) nucleocapsid protein (NP) gene,
complete cds. AY651514 Influenza A virus (A/Ck/HK/YU22/2002(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651521 Influenza A
virus (A/Ck/HK/3176.3/2002(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651522 Influenza A virus (A/Ck/HK/3169.1/2002(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651524 Influenza A
virus (A/feral pigeon/HK/862.7/2002(H5N1)) nucleocapsid protein
(NP) gene, partial cds. AY651525 Influenza A virus (A/tree
sparrow/HK/864/2002(H5N1)) nucleocapsid protein (NP) gene, partial
cds. AY651526 Influenza A virus (A/grey heron/HK/861.1/2002(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651527 Influenza A
virus (A/teal/China/2978.1/2002(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY651523 Influenza A virus (A/black headed
gull/HK/12.1/2003(H5N1)) nucleocapsid protein (NP) gene, partial
cds. AY651487 Influenza A virus (A/Ck/Indonesia/PA/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651515 Influenza A
virus (A/Ck/HK/2133.1/2003(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651516 Influenza A virus (A/Ck/HK/NT93/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651517 Influenza A
virus (A/Ck/HK/SSP141/2003(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651518 Influenza A virus (A/Ck/HK/WF157/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651519 Influenza A
virus (A/Ck/HK/FY157/2003(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651520 Influenza A virus (A/Ck/HK/YU324/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651490 Influenza A
virus (A/Ck/Indonesia/2A/2003(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY676038 Influenza A virus (A/egret/Hong
Kong/757.2/03(H5N1)) nucleoprotein (NP) gene, complete cds.
AY676039 Influenza A virus (A/chicken/Korea/ES/03(H5N1))
nucleoprotein (NP) gene, complete cds. AY676040 Influenza A virus
(A/duck/Korea/ESD1/03(H5N1)) nucleoprotein (NP) gene, complete cds.
AY651529 Influenza A virus (A/Dk/HN/5806/2003(H5N1)) nucleocapsid
protein (NP) gene, partial cds. AY651532 Influenza A virus
(A/Ck/ST/4231/2003(H5N1)) nucleocapsid protein (NP) gene, partial
cds. AY651534 Influenza A virus (A/Dk/ST/4003/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651535 Influenza A
virus (A/Dk/YN/6255/2003(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651536 Influenza A virus (A/Dk/YN/6445/2003(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651485 Influenza A
virus (A/Ck/Indonesia/BL/2003(H5N1)) nucleocapsid protein (NP)
gene, partial cds. AY518364 Influenza A virus
(A/duck/China/E319-2/03(H5N1)) nucleocapsid protein (NP) gene,
complete cds. AY574189 Influenza A virus
(A/chicken/Vietnam/HD1/2004(H5N1)) nucleoprotein gene, partial cds.
AY574192 Influenza A virus (A/chicken/Vietnam/HD2/2004(H5N1))
nucleoprotein gene, partial cds. AJ867076 Influenza A virus
(A/Hatay/2004/(H5N1)) NP gene for nucleoprotein, genomic RNA
AY651486 Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY590579 Influenza A
virus (A/chicken/Nakorn-Patom/Thailand/CU- K2/2004(H5N1))
nucleocapsid protein (NP) gene, complete cds. AY609313 Influenza A
virus (A/chicken/Guangdong/174/04(H5N1)) segment 5, complete
sequence. AY576929 Influenza A virus
(A/chicken/Vietnam/CM/2004(H5N1)) segment 5 nucleoprotein gene,
partial cds. AY576931 Influenza A virus (A/muscovy
duck/Vietnam/MdGL/2004(H5N1)) segment 5 nucleoprotein gene, partial
cds. AB166863 Influenza A virus (A/chicken/Yamaguchi/7/2004(H5N1))
NP gene for nucleoprotein, complete cds. AB188817 Influenza A virus
(A/chicken/Oita/8/2004(H5N1)) NP gene for nucleoprotein, complete
cds. AY651537 Influenza A virus (A/Ck/YN/374/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651538 Influenza A
virus (A/Ck/YN/115/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY653196 Influenza A virus
(A/chicken/Jilin/9/2004(H5N1)) segment 5, complete sequence.
AY651533 Influenza A virus (A/Ph/ST/44/2004(H5N1)) nucleocapsid
protein (NP) gene, partial cds. AY651530 Influenza A virus
(A/Dk/HN/303/2004(H5N1)) nucleocapsid protein (NP) gene, partial
cds. AY651531 Influenza A virus (A/Dk/HN/101/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY684707 Influenza A
virus (A/chicken/Hubei/327/2004(H5N1)) nucleoprotein (NP) gene,
complete cds. AY737290 Influenza A virus
(A/chicken/Guangdong/191/04(H5N1)) segment 5, complete sequence.
AY737297 Influenza A virus (A/chicken/Guangdong/178/04(H5N1))
segment 5, complete sequence. AY737305 Influenza A virus
(A/duck/Guangdong/173/04(H5N1)) segment 5, complete sequence.
AY770081 Influenza A virus (A/chicken/Hubei/489/2004(H5N1))
nucleoprotein (NP) gene, complete cds. AY770996 Influenza A virus
(A/chicken/Ayutthaya/Thailand/CU- 23/04(H5N1)) nucleoprotein gene,
partial cds. AY818139 Influenza A virus
(A/chicken/Vietnam/C58/04(H5N1)) nucleoprotein NP gene, complete
cds. AY818140 Influenza A virus (A/quail/Vietnam/36/04(H5N1))
nucleoprotein NP gene, complete cds. AY856864 Influenza A virus
(A/duck/Shandong/093/2004(H5N1)) segment 5, complete sequence.
AB189046 Influenza A virus (A/chicken/Kyoto/3/2004(H5N1)) NP gene
for nucleoprotein, complete cds,. AB189054 Influenza A virus
(A/crow/Kyoto/53/2004(H5N1)) NP gene for nucleoprotein, complete
cds,. AB189062 Influenza A virus (A/crow/Osaka/102/2004(H5N1)) NP
gene for nucleoprotein, complete cds,. AY651491 Influenza A virus
(A/Ck/Thailand/1/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651492 Influenza A virus
(A/Ck/Thailand/73/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651493 Influenza A virus
(A/Ck/Thailand/9.1/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651494 Influenza A virus
(A/Qa/Thailand/57/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651495 Influenza A virus
(A/bird/Thailand/3.1/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651496 Influenza A virus
(A/Dk/Thailand/71.1/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651497 Influenza A virus
(A/Gs/Thailand/79/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651502 Influenza A virus (A/Ck/Viet
Nam/33/2004(H5N1)) nucleocapsid protein (NP) gene, partial cds.
AY651503 Influenza A virus (A/Ck/Viet Nam/35/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651504 Influenza A
virus (A/Ck/Viet Nam/36/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651505 Influenza A virus (A/Ck/Viet
Nam/37/2004(H5N1)) nucleocapsid protein (NP) gene, partial cds.
AY651506 Influenza A virus (A/Ck/Viet Nam/38/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651507 Influenza A
virus (A/Ck/Viet Nam/39/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651508 Influenza A virus (A/Ck/Viet
Nam/C57/2004(H5N1)) nucleocapsid protein (NP) gene, partial cds.
AY651509 Influenza A virus (A/Dk/Viet Nam/11/2004(H5N1))
nucleocapsid protein (NP) gene, partial cds. AY651488 Influenza A
virus (A/Ck/Indonesia/4/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651489 Influenza A virus
(A/Ck/Indonesia/5/2004(H5N1)) nucleocapsid protein (NP) gene,
partial cds. AY651528 Influenza A virus (A/peregrine
falcon/HK/D0028/2004(H5N1)) nucleocapsid protein (NP) gene, partial
cds. M22573 Influenza A/duck/Hong Kong/7/75 (H3N2), nucleoprotein
(seg 5), RNA. AY180555 Influenza A virus
(A/Chicken/Nanchang/3-120/2001(H3N2)) nucleoprotein (NP) gene,
partial cds. AY779261 Influenza A virus (A/turkey/North
Carolina/12344/03(H3N2)) nucleoprotein (NP) gene, partial cds.
AY779262 Influenza A virus (A/turkey/Minnesota/764-2/03(H3N2))
nucleoprotein (NP) gene, partial cds. AY862655 Influenza A virus
(A/chicken/Korea/S6/03(H3N2)) nucleocapsid protein (NP) gene,
partial cds. AY862656 Influenza A virus (A/duck/Korea/S7/03(H3N2))
nucleocapsid protein (NP) gene, partial cds. AY862657 Influenza A
virus (A/duck/Korea/S8/03(H3N2)) nucleocapsid protein (NP) gene,
partial cds. AY862658 Influenza A virus (A/duck/Korea/S9/03(H3N2))
nucleocapsid protein (NP) gene, partial cds. AY862659 Influenza A
virus (A/duck/Korea/S10/03(H3N2)) nucleocapsid protein (NP) gene,
partial cds. AY862660 Influenza A virus (A/dove/Korea/S11/03(H3N2))
nucleocapsid protein (NP) gene, partial cds. D00050 Influenza A
virus gene for nucleoprotein, complete cds. M14921 Influenza
A/Mallard/NY/6750/78 (H2N2) nucleoprotein (seg 5) RNA, complete
cds. AY422026 Influenza A virus (A/duck/Hokkaido/95/01(H2N2))
nucleoprotein (NP) gene, partial cds. U49093 Influenza A virus
nucleoprotein (NP) mRNA, partial cds. M22574 Influenza
A/duck/Bavaria/2/77 (H1N1), nucleoprotein (seg 5), RNA. M76603
Influenza A/turkey/England/647/77 (H1N1) mRNA, complete cds. M63783
Influenza A/Duck/Australia/749/80 (H1N1) nucleoprotein mRNA,
complete cds. M63778 Influenza A/Turkey/Minnesota/1661/81 (H1N1)
nucleoprotein mRNA, complete cds. Z26855 Influenza virus type A NP
gene for nucleoprotein M76609 Influenza A/turkey/North
Carolina/1790/88 (H1N1) mRNA, complete cds. Z26857 Influenza virus
type A NP gene for nucleoprotein AF213905 Influenza A virus
(A/Mallard/Italy/24/95(H1N1)) segment 5 nucleoprotein (NP) gene,
partial cds. AF213906 Influenza A virus
(A/Chicken/Italy/24/95(H1N1)) segment 5 nucleoprotein (NP) gene,
partial cds. AY633215 Influenza A virus
(A/mallard/Alberta/211/98(H1N1)) nucleoprotein (NP) gene, complete
cds. AY180543 Influenza A virus
(A/Quail/Nanchang/12-340/2000(H1N1)) nucleoprotein (NP) gene,
partial cds. Sequences used in analysis of Influenza A Polymerase
Basic protein 1 (PB1)
AY633218 Influenza A virus (A/mallard/Alberta/211/98(H1N1)) RNA-
directed RNA polymerase subunit P1 (PB1) gene, partial cds. U48284
Influenza A virus polymerase (PB1) mRNA, partial cds. AY180855
Influenza A virus strain A/Quail/Nanchang/12-340/2000 (H1N1)
polymerase subunit PB1 (PB1) gene, partial cds. AY422038 Influenza
A virus (A/duck/Hokkaido/95/01(H2N2)) polymerase subunit (PB1)
gene, partial cds. M25926 Influenza A/Mallard/New York/6750/78
(H2N2) PB1 gene, complete cds. AY180871 Influenza A virus strain
A/Chicken/Nanchang/3-120/2001 (H3N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY779265 Influenza A virus (A/turkey/North
Carolina/12344/03(H3N2)) polymerase basic protein 1 (PB1) gene,
partial cds. AY779266 Influenza A virus
(A/turkey/Minnesota/764-2/03(H3N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY862703 Influenza A virus
(A/chicken/Korea/S6/03(H3N2)) PB1 (PB1) gene, partial cds. AY862704
Influenza A virus (A/duck/Korea/S7/03(H3N2)) PB1 (PB1) gene,
partial cds. AY862705 Influenza A virus (A/duck/Korea/S8/03(H3N2))
PB1 (PB1) gene, partial cds. AY862706 Influenza A virus
(A/duck/Korea/S9/03(H3N2)) PB1 (PB1) gene, partial cds. AY862707
Influenza A virus (A/duck/Korea/S10/03(H3N2)) PB1 (PB1) gene,
partial cds. AY862708 Influenza A virus (A/dove/Korea/S11/03(H3N2))
PB1 (PB1) gene, partial cds. AF213911 Influenza A virus
(A/Chicken/Italy/5945/95(H3N2)) segment 8 PB1 polymerase protein
gene, partial cds. AB166860 Influenza A virus
(A/chicken/Yamaguchi/7/2004(H5N1)) PB1 gene for polymerase basic
protein 1, complete cds. AB188814 Influenza A virus
(A/chicken/Oita/8/2004(H5N1)) PB1 gene for polymerase basic protein
1, complete cds. AF398423 Influenza A virus (A/Goose/Hong
Kong/385.3/2000(H5N1)) polymerase (PB1) gene, partial cds. AF398424
Influenza A virus (A/Goose/Hong Kong/385.5/2000(H5N1)) polymerase
(PB1) gene, partial cds. AF468839 Influenza A virus
(A/Duck/Anyang/AVL-1/2001(H5N1)) polymerase basic protein 1 (PB1)
gene, complete cds. AF509169 Influenza A virus (A/Chicken/Hong
Kong/FY77/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509170
Influenza A virus (A/Chicken/Hong Kong/YU562/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509171 Influenza A virus (A/Chicken/Hong
Kong/YU563/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509172
Influenza A virus (A/Chicken/Hong Kong/FY150/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509173 Influenza A virus
(A/Pheasant/Hong Kong/FY155/01 (H5N1)) polymerase (PB1) gene,
partial cds. AF509174 Influenza A virus (A/Silky Chicken/Hong
Kong/SF189/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509175
Influenza A virus (A/Quail/Hong Kong/SF203/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509176 Influenza A virus (A/Pigeon/Hong
Kong/SF215/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509177
Influenza A virus (A/Chicken/Hong Kong/SF219/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509178 Influenza A virus (A/Chicken/Hong
Kong/715.5/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509179
Influenza A virus (A/Chicken/Hong Kong/751.1/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509180 Influenza A virus (A/Chicken/Hong
Kong/822.1/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509181
Influenza A virus (A/Chicken/Hong Kong/829.2/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509182 Influenza A virus (A/Chicken/Hong
Kong/830.2/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509183
Influenza A virus (A/Chicken/Hong Kong/858.3/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509184 Influenza A virus (A/Chicken/Hong
Kong/866.3/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509185
Influenza A virus (A/Chicken/Hong Kong/867.1/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509186 Influenza A virus (A/Chicken/Hong
Kong/879.1/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509187
Influenza A virus (A/Chicken/Hong Kong/873.3/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509188 Influenza A virus (A/Chicken/Hong
Kong/876.1/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509189
Influenza A virus (A/Chicken/Hong Kong/891.1/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509190 Influenza A virus (A/Chicken/Hong
Kong/893.2/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509191
Influenza A virus (A/Goose/Hong Kong/76.1/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509192 Influenza A virus (A/Goose/Hong
Kong/ww100/01 (H5N1)) polymerase (PB1) gene, partial cds. AF509193
Influenza A virus (A/Duck/Hong Kong/573.4/01 (H5N1)) polymerase
(PB1) gene, partial cds. AF509194 Influenza A virus (A/Duck/Hong
Kong/646.3/01 (H5N1)) polymerase (PB1) gene, partial cds. AY035888
Influenza A virus (A/goose/Guangdong/3/97(H5N1)) polymerase basic
protein 1 (PB1) gene, complete cds. AY059513 Influenza A virus
(A/Goose/Hong Kong/ww26/2000(H5N1)) segment 2 polymerase (PB1)
gene, partial cds. AY059514 Influenza A virus (A/Goose/Hong
Kong/ww28/2000(H5N1)) segment 2 polymerase (PB1) gene, partial cds.
AY059515 Influenza A virus (A/Duck/Hong Kong/ww381/2000(H5N1))
segment 2 polymerase (PB1) gene, partial cds. AY059516 Influenza A
virus (A/Duck/Hong Kong/ww461/2000(H5N1)) segment 2 polymerase
(PB1) gene, partial cds. AY059517 Influenza A virus (A/Goose/Hong
Kong/ww491/2000(H5N1)) segment 2 polymerase (PB1) gene, partial
cds. AY059518 Influenza A virus (A/Duck/Hong
Kong/2986.1/2000(H5N1)) segment 2 polymerase (PB1) gene, partial
cds. AY059519 Influenza A virus (A/Goose/Hong
Kong/3014.8/2000(H5N1)) segment 2 polymerase (PB1) gene, partial
cds. AY221575 Influenza A virus
(A/Chicken/HongKong/NT873.3/01-MB(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221576 Influenza A virus
(A/Chicken/HongKong/NT873.3/01(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221577 Influenza A virus
(A/Chicken/HongKong/FY150/01-MB(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221578 Influenza A virus
(A/Chicken/HongKong/FY150/01(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221579 Influenza A virus
(A/Pheasant/HongKong/FY155/01-MB(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221580 Influenza A virus
(A/Pheasant/HongKong/FY155/01(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221581 Influenza A virus
(A/Chicken/HongKong/YU822.2/01-MB(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221582 Influenza A virus
(A/Chicken/HongKong/YU822.2/01(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY221583 Influenza A virus
(A/Chicken/HongKong/YU562/01(H5N1)) polymerase basic protein 1
(PB1) gene, partial cds. AY518366 Influenza A virus
(A/duck/China/E319-2/03(H5N1)) polymerase subunit PB1 (PB1) gene,
complete cds. AY576394 Influenza A virus (A/Gs/HK/739.2/02 (H5N1))
polymerase (PB1) gene, partial cds. AY576395 Influenza A virus
(A/Eg/HK/757.3/02 (H5N1)) polymerase (PB1) gene, partial cds.
AY576396 Influenza A virus (A/G.H/HK/793.1/02 (H5N1)) polymerase
(PB1) gene, partial cds. AY576397 Influenza A virus (A/Dk/HK/821/02
(H5N1)) polymerase (PB1) gene, complete cds. AY576398 Influenza A
virus (A/Ck/HK/31.4/02 (H5N1)) polymerase (PB1) gene, partial cds.
AY576399 Influenza A virus (A/Ck/HK/61.9/02 (H5N1)) polymerase
(PB1) gene, partial cds. AY576400 Influenza A virus
(A/Ck/HK/YU777/02 (H5N1)) polymerase (PB1) gene, partial cds.
AY576401 Influenza A virus (A/Ck/HK/96.1/02 (H5N1)) polymerase
(PB1) gene, partial cds. AY576402 Influenza A virus
(A/Ck/HK/409.1/02 (H5N1)) polymerase (PB1) gene, partial cds.
AY576403 Influenza A virus (A/Ph/HK/674.15/02 (H5N1)) polymerase
(PB1) gene, partial cds. AY585483 Influenza A virus
(A/duck/Fujian/01/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585484 Influenza A virus
(A/duck/Fujian/13/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585485 Influenza A virus
(A/duck/Fujian/17/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585486 Influenza A virus
(A/duck/Fujian/19/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585487 Influenza A virus
(A/duck/Guangdong/01/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585488 Influenza A virus
(A/duck/Guangdong/07/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585489 Influenza A virus
(A/duck/Guangdong/12/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585490 Influenza A virus
(A/duck/Guangdong/22/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585491 Influenza A virus
(A/duck/Guangdong/40/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585492 Influenza A virus
(A/duck/Guangxi/07/1999(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585493 Influenza A virus
(A/duck/Guangxi/22/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585494 Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585495 Influenza A virus
(A/duck/Guangxi/50/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585496 Influenza A virus
(A/duck/Guangxi/53/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585497 Influenza A virus
(A/duck/Shanghai/08/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585498 Influenza A virus
(A/duck/Shanghai/13/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585499 Influenza A virus
(A/duck/Shanghai/35/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585500 Influenza A virus
(A/duck/Shanghai/37/2002(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585501 Influenza A virus
(A/duck/Shanghai/38/2001(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585502 Influenza A virus
(A/duck/Zhejiang/11/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY585503 Influenza A virus
(A/duck/Zhejiang/52/2000(H5N1)) polymerase basic protein 1 (PB1)
mRNA, complete cds. AY590582 Influenza A virus
(A/chicken/Nakorn-Patom/Thailand/CU- K2/2004(H5N1)) polymerase
basic protein 1 (PBP1) gene, complete cds. AY609310 Influenza A
virus (A/chicken/Guangdong/174/04 (H5N1)) segment 2, complete
sequence. AY651651 Influenza A virus (A/Ck/Indonesia/BL/2003(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651652
Influenza A virus (A/Dk/Indonesia/MS/2004(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651653 Influenza A virus
(A/Ck/Indonesia/PA/2003(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651654 Influenza A virus
(A/Ck/Indonesia/4/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651655 Influenza A virus
(A/Ck/Indonesia/2A/2003(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651656 Influenza A virus
(A/Ck/Indonesia/5/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651657 Influenza A virus
(A/Ck/Thailand/1/2004(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651658 Influenza A virus
(A/Ck/Thailand/73/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651659 Influenza A virus
(A/Ck/Thailand/9.1/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651660 Influenza A virus
(A/Qa/Thailand/57/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651661 Influenza A virus
(A/bird/Thailand/3.1/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651662 Influenza A virus
(A/Dk/Thailand/71.1/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651663 Influenza A virus
(A/Gs/Thailand/79/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651668 Influenza A virus (A/Ck/Viet
Nam/33/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial
cds. AY651669 Influenza A virus (A/Ck/Viet Nam/35/2004(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651670
Influenza A virus (A/Ck/Viet Nam/36/2004(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651671 Influenza A virus
(A/Ck/Viet Nam/37/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651672 Influenza A virus (A/Ck/Viet
Nam/38/2004(H5N1)) polymerase basic subunit 1 (PB1) gene, partial
cds. AY651673 Influenza A virus (A/Ck/Viet Nam/39/2004(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651674
Influenza A virus (A/Ck/Viet Nam/C57/2004(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651675 Influenza A virus
(A/Dk/Viet Nam/11/2004(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651676 Influenza A virus
(A/Gf/HK/38/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651677 Influenza A virus (A/Ck/HK/31.2/2002(H5N1))
polymerase basic subunit 1 gene, partial cds. AY651678 Influenza A
virus (A/Ck/HK/37.4/2002(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651679 Influenza A virus
(A/SCk/HK/YU100/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651680 Influenza A virus (A/Ck/HK/YU22/2002(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651681
Influenza A virus (A/Ck/HK/3176.3/2002(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651682 Influenza A virus
(A/Ck/HK/3169.1/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651683 Influenza A virus (A/Ck/HK/FY157/2003(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds.
AY651684 Influenza A virus (A/Ck/HK/YU324/2003(H5N1)) polymerase
basic subunit 1 (PB1) gene, partial cds. AY651685 Influenza A virus
(A/Ck/HK/2133.1/2003(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651686 Influenza A virus (A/Ck/HK/NT93/2003(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651687
Influenza A virus (A/Ck/HK/SSP141/2003(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651688 Influenza A virus
(A/Ck/HK/WF157/2003(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651689 Influenza A virus (A/black headed
gull/HK/12.1/2003(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651690 Influenza A virus (A/feral
pigeon/HK/862.7/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651691 Influenza A virus (A/grey
heron/HK/861.1/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651692 Influenza A virus (A/tree
sparrow/HK/864/2002(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651693 Influenza A virus
(A/teal/China/2978.1/2002(H5N1)) polymerase basic subunit 1 (PB1)
gene, partial cds. AY651694 Influenza A virus (A/peregrine
falcon/HK/D0028/2004(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651695 Influenza A virus (A/Dk/HN/5806/2003(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651696
Influenza A virus (A/Dk/ST/4003/2003(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651697 Influenza A virus
(A/Ck/ST/4231/2003(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651698 Influenza A virus (A/Dk/YN/6255/2003(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651699
Influenza A virus (A/Dk/YN/6445/2003(H5N1)) polymerase basic
subunit 1 (PB1) gene, partial cds. AY651700 Influenza A virus
(A/Ph/ST/44/2004(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651701 Influenza A virus (A/Dk/HN/303/2004(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY651702
Influenza A virus (A/Dk/HN/101/2004(H5N1)) polymerase basic subunit
1 (PB1) gene, partial cds. AY651703 Influenza A virus
(A/Ck/YN/374/2004(H5N1)) polymerase basic subunit 1 (PB1) gene,
partial cds. AY651704 Influenza A virus (A/Ck/YN/115/2004(H5N1))
polymerase basic subunit 1 (PB1) gene, partial cds. AY653199
Influenza A virus (A/chicken/Jilin/9/2004(H5N1)) segment 2,
complete sequence. AY676025 Influenza A virus strain (A/duck/Hong
Kong/821/02(H5N1)) polymerase basic 1 (PB1) gene, complete cds.
AY676026 Influenza A virus strain (A/egret/Hong
Kong/757.2/03(H5N1)) polymerase basic 1 (PB1) gene, complete cds.
AY676027 Influenza A virus strain (A/chicken/Korea/ES/03(H5N1))
polymerase basic 1 (PB1) gene, complete cds. AY676028 Influenza A
virus strain (A/duck/Korea/ESD1/03(H5N1)) polymerase basic 1 (PB1)
gene, complete cds. AY684704 Influenza A virus
(A/chicken/Hubei/327/2004(H5N1)) polymerase basic protein 1 (PB1)
gene, complete cds. AY737287 Influenza A virus
(A/chicken/Guangdong/191/04(H5N1)) segment 2, complete sequence.
AY737294 Influenza A virus (A/chicken/Guangdong/178/04(H5N1))
segment 2, complete sequence. AY737302 Influenza A virus
(A/duck/Guangdong/173/04(H5N1)) segment 2, complete sequence.
AY770083 Influenza A virus (A/chicken/Hubei/489/2004(H5N1))
nonfunctional polymerase basic protein 1 (PB1) gene, complete
sequence. AY770994 Influenza A virus
(A/chicken/Ayutthaya/Thailand/CU- 23/04(H5N1)) polymerase basic
protein 1 gene, partial cds. AY818130 Influenza A virus
(A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PB1 gene,
complete cds. AY818131 Influenza A virus
(A/quail/Vietnam/36/04(H5N1)) polymerase protein PB1 gene, complete
cds. AY856862 Influenza A virus (A/duck/Shandong/093/2004(H5N1))
segment 2, complete sequence. AB188822 Influenza A virus
(A/chicken/Kyoto/3/2004(H5N1)) PB1 gene for polymerase basic
protein 1, complete cds. AB189051 Influenza A virus
(A/crow/Kyoto/53/2004(H5N1)) PB1 gene for polymerase basic protein
1, complete cds,. AB189060 Influenza A virus
(A/crow/Osaka/102/2004(H5N1)) PB1 gene for polymerase basic protein
1, complete cds,. AF046085 Influenza A virus (A/Chicken/Hong
Kong/220/97 (H5N1)) polymerase basic protein 1 (PB1) gene, complete
cds. AF098590 Influenza A virus (A/Chicken/Hong Kong/258/97 (H5N1))
PB1 protein (PB1) gene, partial cds. AF098591 Influenza A virus
(A/Chicken/Hong Kong/y388/97 (H5N1)) PB1 protein (PB1) gene,
partial cds. AF098592 Influenza A virus (A/Chicken/Hong Kong/728/97
(H5N1)) PB1 protein (PB1) gene, partial cds. AF098593 Influenza A
virus (A/Chicken/Hong Kong/786/97 (H5N1)) PB1 protein (PB1) gene,
partial cds. AF098594 Influenza A virus (A/Chicken/Hong Kong/915/97
(H5N1)) PB1 protein (PB1) gene, partial cds. AF098595 Influenza A
virus (A/Duck/Hong Kong/p46/97 (H5N1)) PB1 protein (PB1) gene,
partial cds. AF098596 Influenza A virus (A/Duck/Hong Kong/y283/97
(H5N1)) PB1 protein (PB1) gene, partial cds. AF098598 Influenza A
virus (A/Goose/Hong Kong/w355/97 (H5N1)) PB1 protein (PB1) gene,
partial cds. AF144301 Influenza A virus
(A/Goose/Guangdong/1/96(H5N1)) polymerase (PB1) gene, complete cds.
AF216716 Influenza A virus (A/Environment/Hong Kong/437-4/99
(H5N1)) polymerase basic protein 1 gene, complete cds. AF216724
Influenza A virus (A/Environment/Hong Kong/437-6/99 (H5N1))
polymerase basic protein 1 gene, complete cds. AF216732 Influenza A
virus (A/Environment/Hong Kong/437-8/99 (H5N1)) polymerase basic
protein 1 gene, complete cds. AF216740 Influenza A virus
(A/Environment/Hong Kong/437-10/99 (H5N1)) polymerase basic protein
1 gene, complete cds. AY303663 Influenza A virus
(A/chicken/Chile/176822/02(H7N3)) polymerase basic protein 1 gene,
complete cds. AY303664 Influenza A virus
(A/chicken/Chile/4957/02(H7N3)) polymerase basic protein 1 gene,
partial cds. AY586435 Influenza A Virus
(A/turkey/Italy/214845/02(H7N3)) PB1 gene, partial cds. AY586436
Influenza A virus (A/turkey/Italy/220158/2002(H7N3)) PB1 gene,
partial cds. AY586437 Influenza A virus
(A/mallard/Italy/33/01(H7N3)) PB1 gene, partial cds. AY586438
Influenza A Virus (A/mallard/Italy/43/01(H7N3)) PB1 gene, partial
cds. AY616765 Influenza A virus (A/chicken/British
Columbia/04(H7N3)) PB1 polymerase subunit (PB1) gene, complete cds.
AY646084 Influenza A virus (A/chicken/British
Columbia/GSC_human_B/04(H7N3)) polymerase basic protein 1 (PB1)
gene, complete cds. AY648293 Influenza A virus
(A/GSC_chicken_B/British Columbia/04(H7N3)) PB1 polymerase subunit
(PB1) gene, complete cds. AY653039 Influenza A virus
(A/GSC_chicken/British Columbia/04(H7N3)) PB1 polymerase subunit
(PB1) gene, complete cds. AJ620348 Influenza A virus
(A/Chicken/Germany/R28/03(H7N7)) PB1 gene for RNA polymerase,
genomic RNA. AY340080 Influenza A virus
(A/Netherlands/124/03(H7N7)) polymerase (PB1) gene, partial cds.
AY340081 Influenza A virus (A/Netherlands/126/03(H7N7)) polymerase
(PB1) gene, partial cds. AY340082 Influenza A virus
(A/Netherlands/127/03(H7N7)) polymerase (PB1) gene, partial cds.
AY340083 Influenza A virus (A/Netherlands/219/03(H7N7)) polymerase
(PB1) gene, complete cds. AY340084 Influenza A virus
(A/Netherlands/033/03(H7N7)) polymerase (PB1) gene, complete cds.
AY340085 Influenza A virus (A/chicken/Netherlands/1/03(H7N7))
polymerase (PB1) gene, complete cds. AB049155 Influenza A virus
(A/parakeet/Chiba/1/97(H9N2)) PB1 gene for polymerase basic protein
1, complete cds. AB049156 Influenza A virus
(A/parakeet/Narita/92A/98(H9N2)) PB1 gene for polymerase basic
protein 1, complete cds. AF508618 Influenza A virus
(A/Ostrich/South Africa/9508103/95(H9N2)) segment 2 polymerase PB1
(PB1) gene, partial cds. AF508619 Influenza A virus
(A/Chicken/Pakistan/4/99(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508620 Influenza A virus
(A/Chicken/Pakistan/5/99(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508621 Influenza A virus
(A/Chicken/Germany/R45/98(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508622 Influenza A virus
(A/Duck/Germany/113/95(H9N2)) segment 2 polymerase PB1 (PB1) gene,
partial cds. AF508623 Influenza A virus
(A/Chicken/Iran/11T/99(H9N2)) segment 2 polymerase PB1 (PB1) gene,
partial cds. AF508624 Influenza A virus (A/Chicken/Saudi
Arabia/532/99(H9N2)) segment 2 polymerase PB1 (PB1) gene, partial
cds. AF508625 Influenza A virus (A/Pheasant/Ireland/PV18/97(H9N2))
segment 2 polymerase PB1 (PB1) gene, partial cds. AF508626
Influenza A virus (A/Chicken/Korea/99029/99(H9N2)) segment 2
polymerase PB1 (PB1) gene, partial cds. AF508627 Influenza A virus
(A/Chicken/Beijing/8/98(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF508628 Influenza A virus
(A/Chicken/Guangdong/10/00(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508629 Influenza A virus
(A/Chicken/Guangdong/11/97(H9N2)) segment 2 polymerase PB1 (PB1)
gene, complete cds. AF508630 Influenza A virus
(A/Chicken/Hebei/4/98(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF508631 Influenza A virus
(A/Chicken/Heilongjiang/10/97(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508632 Influenza A virus
(A/Chicken/Henan/62/00(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF508633 Influenza A virus
(A/Chicken/Ningxia/5/99(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF508634 Influenza A virus
(A/Chicken/Sichuan/5/97(H9N2)) segment 2 polymerase PB1 (PB1) gene,
partial cds. AF508635 Influenza A virus
(A/Chicken/Shandong/6/96(H9N2)) segment 2 polymerase PB1 (PB1)
gene, partial cds. AF508636 Influenza A virus
(A/Chicken/Shijiazhuang/2/99(H9N2)) segment 2 polymerase PB1 (PB1)
gene, complete cds. AF508637 Influenza A virus
(A/Chicken/Shenzhen/9/97(H9N2)) segment 2 polymerase PB1 (PB1)
gene, complete cds. AF508638 Influenza A virus
(A/Duck/Nanjing/1/97(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF508639 Influenza A virus
(A/Quail/Shanghai/8/96(H9N2)) segment 2 polymerase PB1 (PB1) gene,
complete cds. AF523427 Influenza A virus
(A/Duck/Shantou/830/00(H9N2)) polymerase (PB1) gene, partial cds.
AF523428 Influenza A virus (A/Duck/Shantou/2102/00(H9N2))
polymerase (PB1) gene, partial cds. AF523429 Influenza A virus
(A/Duck/Shantou/1043/00(H9N2)) polymerase (PB1) gene, partial cds.
AF523430 Influenza A virus (A/Duck/Shantou/2134/00(H9N2))
polymerase (PB1) gene, partial cds. AF523431 Influenza A virus
(A/Wild Duck/Shantou/4808/01(H9N2)) polymerase (PB1) gene, partial
cds. AF523432 Influenza A virus (A/Duck/Shantou/2144/00(H9N2))
polymerase (PB1) gene, partial cds. AF523433 Influenza A virus
(A/Duck/Shantou/2143/00(H9N2)) polymerase (PB1) gene, partial cds.
AF523434 Influenza A virus (A/Duck/Shantou/1796/00(H9N2))
polymerase (PB1) gene, partial cds. AF523435 Influenza A virus
(A/Duck/Shantou/2088/01(H9N2)) polymerase (PB1) gene, partial cds.
AF523436 Influenza A virus (A/Duck/Shantou/1881/00(H9N2))
polymerase (PB1) gene, partial cds. AF523437 Influenza A virus
(A/Duck/Hong Kong/366/78(H9N2)) polymerase (PB1) gene, partial cds.
AF523438 Influenza A virus (A/Duck/Hong Kong/552/79(H9N2))
polymerase (PB1) gene, partial cds. AF523439 Influenza A virus
(A/Duck/Hong Kong/86/76(H9N2)) polymerase (PB1) gene, partial cds.
AF523440 Influenza A virus (A/Duck/Hong Kong/289/78(H9N2))
polymerase (PB1) gene, partial cds. AF523441 Influenza A virus
(A/Duck/Hong Kong/610/79(H9N2)) polymerase (PB1) gene, partial cds.
AF523442 Influenza A virus (A/Duck/Shantou/1605/01(H9N2))
polymerase (PB1) gene, partial cds. AF523443 Influenza A virus
(A/Duck/Shantou/1042/00(H9N2)) polymerase (PB1) gene, partial cds.
AF536659 Influenza A virus (A/Chicken/Beijing/1/95(H9N2)) PB1 gene,
partial cds. AF536660 Influenza A virus
(A/Chicken/Beijing/2/97(H9N2)) PB1 gene, partial cds. AF536661
Influenza A virus (A/Chicken/Beijing/3/99(H9N2)) PB1 gene, partial
cds. AF536662 Influenza A virus (A/Chicken/Guangdong/97(H9N2)) PB1
gene, partial cds. AF536663 Influenza A virus
(A/Chicken/Hebei/1/96(H9N2)) PB1 gene, partial cds. AF536664
Influenza A virus (A/Chicken/Hebei/2/98(H9N2)) PB1 gene, partial
cds. AF536665 Influenza A virus (A/Chicken/Hebei/3/98(H9N2)) PB1
gene, partial cds. AF536666 Influenza A virus
(A/Chicken/Henan/98(H9N2)) PB1 gene, partial cds. AF536667
Influenza A virus (A/Chicken/Liaoning/99(H9N2)) PB1 gene, partial
cds. AF536668 Influenza A virus (A/Chicken/Shandong/98(H9N2)) PB1
gene, partial cds. AJ291396 Influenza A virus
(A/Chicken/Pakistan/2/99 (H9N2)) PB1 gene for polymerase PB1,
genomic RNA. AJ427862 Influenza A virus (A/quail/Hong Kong/FY298/00
(H9N2)) partial pb1 gene for PB1 polymerase protein, genomic RNA
AY180840 Influenza A virus strain A/Pigeon/Nanchang/7-058/2000
(H9N2) polymerase subunit PB1 (PB1) gene, partial cds. AY180843
Influenza A virus strain A/Quail/Nanchang/2-0460/2000 (H9N2)
polymerase subunit PB1 (PB1) gene, partial cds. AY180844 Influenza
A virus strain A/Pigeon/Nanchang/2-0461/2000 (H9N2) polymerase
subunit PB1 (PB1) gene, partial cds.
AY180851 Influenza A virus strain A/Pigeon/Nanchang/11-145/2000
(H9N2) polymerase subunit PB1 (PB1) gene, partial cds. AY180852
Influenza A virus strain A/Duck/Nanchang/11-197/2000 (H9N2)
polymerase subunit PB1 (PB1) gene, partial cds. AY180854 Influenza
A virus strain A/Duck/Nanchang/11-290/2000 (H9N2) polymerase
subunit PB1 (PB1) gene, partial cds. AY180856 Influenza A virus
strain A/Duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PB1
(PB1) gene, partial cds. AY180866 Influenza A virus strain
A/Duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180867 Influenza A virus strain
A/Chicken/Nanchang/1-0016/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180873 Influenza A virus strain
A/Chicken/Nanchang/4-010/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180874 Influenza A virus strain
A/Chicken/Nanchang/4-301/2001 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180875 Influenza A virus strain
A/Chicken/Nanchang/4-361/2001 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180892 Influenza A virus strain
A/Quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180897 Influenza A virus strain A/Wild
Duck/Nanchang/2-0480/2000 (H9N2) polymerase subunit PB1 (PB1) gene,
partial cds. AY180900 Influenza A virus strain
A/Duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY180901 Influenza A virus strain
A/Duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PB1 (PB1)
gene, partial cds. AY253751 Influenza A virus
(A/Chicken/Shanghai/F/98(H9N2)) polymerase basic protein 1 (PB1)
gene, complete cds. AY307947 Influenza A virus
(A/chicken/Beijing/1/00(H9N2)) polymerase subunit (PB1) gene,
partial cds. AY307948 Influenza A virus
(A/chicken/Hebei/1/01(H9N2)) polymerase subunit (PB1) gene, partial
cds. AY633170 Influenza A virus (A/mallard/Alberta/17/91(H9N2))
RNA-directed RNA polymerase subunit P1 (PB1) gene, partial cds.
AY633282 Influenza A virus (A/mallard/Alberta/321/88(H9N2)) RNA-
directed RNA polymerase subunit P1 (PB1) gene, partial cds.
AY633298 Influenza A virus (A/mallard/Alberta/11/91(H9N2))
RNA-directed RNA polymerase subunit P1 (PB1) gene, partial cds.
AY664774 Influenza A virus (A/chicken/HongKong/CSW153/03(H9N2))
polymerase basic protein 1 (PB1) gene, partial cds. AY664775
Influenza A virus (A/chicken/HongKong/AP45/03(H9N2)) polymerase
basic protein 1 (PB1) gene, partial cds. AY664776 Influenza A virus
(A/chicken/HongKong/BD90/03(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY664777 Influenza A virus
(A/chicken/HongKong/CSW291/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664778 Influenza A virus
(A/chicken/HongKong/CSW304/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664779 Influenza A virus
(A/chicken/HongKong/FY23/03(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY664780 Influenza A virus
(A/guineafowl/HongKong/NT101/03(H9N2) polymerase basic protein 1
(PB1) gene, partial cds. AY664781 Influenza A virus
(A/chicken/HongKong/NT142/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664782 Influenza A virus
(A/chicken/HongKong/SF1/03(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY664783 Influenza A virus
(A/chicken/HongKong/SSP101/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664784 Influenza A virus
(A/chicken/HongKong/TP38/03(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY664785 Influenza A virus
(A/chicken/HongKong/WF126/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664786 Influenza A virus
(A/pigeon/HongKong/WF53/03(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY664787 Influenza A virus
(A/pheasant/HongKong/WF54/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664788 Influenza A virus
(A/guineafowl/HongKong/NT184/03(H9N2) polymerase basic protein 1
(PB1) gene, partial cds. AY664789 Influenza A virus
(A/chicken/HongKong/WF120/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664790 Influenza A virus
(A/chicken/HongKong/NT366/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY664791 Influenza A virus
(A/chicken/HongKong/YU427/03(H9N2)) polymerase basic protein 1
(PB1) gene, partial cds. AY800239 Influenza A virus
(A/chicken/Korea/S1/2003(H9N2)) polymerase basic protein 1 (PB1)
gene, partial cds. AY862694 Influenza A virus (A/silky
chicken/Korea/S3/03(H9N2)) PB1 (PB1) gene, partial cds. AY862695
Influenza A virus (A/chicken/Korea/S4/03(H9N2)) PB1 (PB1) gene,
partial cds. AY862696 Influenza A virus
(A/chicken/Korea/S5/03(H9N2)) PB1 (PB1) gene, partial cds. AY862697
Influenza A virus (A/chicken/Korea/S12/03(H9N2)) PB1 (PB1) gene,
partial cds. AY862698 Influenza A virus (A/duck/Korea/S13/03(H9N2))
PB1 (PB1) gene, partial cds. AY862699 Influenza A virus
(A/dove/Korea/S14/03(H9N2)) PB1 (PB1) gene, partial cds. AY862700
Influenza A virus (A/chicken/Korea/S15/03(H9N2)) PB1 (PB1) gene,
partial cds. AY862701 Influenza A virus
(A/chicken/Korea/S16/03(H9N2)) PB1 (PB1) gene, partial cds.
AY862702 Influenza A virus (A/chicken/Korea/S18/03(H9N2)) PB1 (PB1)
gene, partial cds. AF156416 Influenza A virus (A/Chicken/Hong
Kong/G9/97(H9N2)) segment 2 PB1 polymerase subunit (PB1) gene,
partial cds. AF156417 Influenza A virus (A/Chicken/Hong Kong/G23/99
(H9N2)) segment 2 PB1 polymerase subunit (PB1) gene, partial cds.
AF156418 Influenza A virus (A/Pigeon/Hong Kong/Y233/97(H9N2))
segment 2 PB1 polymerase subunit (PB1) gene, partial cds. AF156419
Influenza A virus (A/Duck/Hong Kong/Y280/97(H9N2)) segment 2 PB1
polymerase subunit (PB1) gene, partial cds. AF156420 Influenza A
virus (A/Duck/Hong Kong/Y439/97(H9N2)) segment 2 PB1 polymerase
subunit (PB1) gene, partial cds. AF156421 Influenza A virus
(A/Quail/Hong Kong/G1/97 (H9N2)) segment 2 PB1 polymerase subunit
(PB1) gene, partial cds. AF156422 Influenza A virus (A/Chicken/Hong
Kong/739/94(H9N2)) segment 2 PB1 polymerase subunit (PB1) gene,
partial cds. AF156423 Influenza A virus
(A/Chicken/Beijing/1/94(H9N2)) segment 2 PB1 polymerase subunit
(PB1) gene, partial cds. AF156424 Influenza A virus (A/Quail/Hong
Kong/AF157/92(H9N2)) segment 2 PB1 polymerase subunit (PB1) gene,
partial cds. AF156425 Influenza A virus
(A/Chicken/Korea/38349-p96323/96(H9N2)) segment 2 PB1 polymerase
subunit (PB1) gene, partial cds. AF156426 Influenza A virus
(A/Chicken/Korea/25232-96006/96(H9N2)) segment 2 PB1 polymerase
subunit (PB1) gene, partial cds. AF156427 Influenza A virus
(A/Shorebird/Delaware/9/96(H9N2)) segment 2 PB1 polymerase subunit
(PB1) gene, partial cds. AF156428 Influenza A virus
(A/Quail/Arkansas/29209-1/93(H9N2)) segment 2 PB1 polymerase
subunit (PB1) gene, partial cds. AF156429 Influenza A virus
(A/Turkey/California/189/66(H9N2)) segment 2 PB1 polymerase subunit
(PB1) gene, partial cds. AF222632 Influenza A virus (A/Quail/Hong
Kong/A17/99(H9N2)) segment 2 polymerase 1 (PB1) gene, partial cds.
AF222633 Influenza A virus (A/Pigeon/Hong Kong/FY6/99(H9N2))
segment 2 polymerase 1 (PB1) gene, partial cds. AF222634 Influenza
A virus (A/Chicken/Hong Kong/NT16/99(H9N2)) segment 2 polymerase 1
(PB1) gene, partial cds. AF222635 Influenza A virus (A/Quail/Hong
Kong/SSP10/99(H9N2)) segment 2 polymerase 1 (PB1) gene, partial
cds. AF222636 Influenza A virus (A/Pheasant/Hong
Kong/SSP11/99(H9N2)) segment 2 polymerase 1 (PB1) gene, partial
cds. AF222637 Influenza A virus (A/Chicken/Hong Kong/FY20/99(H9N2))
segment 2 polymerase 1 (PB1) gene, partial cds. AF222638 Influenza
A virus (A/Chicken/Hong Kong/KC12/99(H9N2)) segment 2 polymerase 1
(PB1) gene, partial cds. AF222639 Influenza A virus (A/Quail/Hong
Kong/NT2899(H9N2)) segment 2 polymerase 1 (PB1) gene, partial cds.
AF222640 Influenza A virus (A/Chicken/Hong Kong/SF2/99(H9N2))
segment 2 polymerase 1 (PB1) gene, partial cds. AF222641 Influenza
A virus (A/Silky Chicken/Hong Kong/SF44/99(H9N2)) segment 2
polymerase 1 (PB1) gene, partial cds. Sequences used in analysis of
Influenza A Polymerase Basic protein 2 (PB2) gi|49356919|AY633219|
Influenza A virus (A/mallard/Alberta/211/98(H1N1)) polymerase basic
protein 2 (PB2) gene, partial cds. gi|27466107|AY180748| Influenza
A virus strain A/Quail/Nanchang/12-340/2000 (H1N1) polymerase
subunit PB2 (PB2) gene, partial cds. gi|45272173|AY422042|
Influenza A virus (A/duck/Hokkaido/95/01(H2N2)) polymerase subunit
(PB2) gene, partial cds. gi|18091825|AF213910| Influenza A virus
(A/Chicken/Italy/5945/95(H3N2)) segment 1 PB2 polymerase protein
gene, partial cds. gi|27466133|AY180761| Influenza A virus strain
A/Chicken/Nanchang/3- 120/2001 (H3N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|56160002|AY779267| Influenza A virus
(A/turkey/North Carolina/12344/03(H3N2)) polymerase basic protein 2
(PB2) gene, partial cds. gi|56160004|AY779268| Influenza A virus
(A/turkey/Minnesota/764-2/03(H3N2)) polymerase basic protein 2
(PB2) gene, partial cds. gi|58429704|AY862719| Influenza A virus
(A/chicken/Korea/S6/03(H3N2)) PB2 (PB2) gene, partial cds.
gi|58429706|AY862720| Influenza A virus (A/duck/Korea/S7/03(H3N2))
PB2 (PB2) gene, partial cds. gi|58429708|AY862721| Influenza A
virus (A/duck/Korea/S8/03(H3N2)) PB2 (PB2) gene, partial cds.
gi|58429710|AY862722| Influenza A virus (A/duck/Korea/S9/03(H3N2))
PB2 (PB2) gene, partial cds. gi|58429712|AY862723| Influenza A
virus (A/duck/Korea/S10/03(H3N2)) PB2 (PB2) gene, partial cds.
gi|58429714|AY862724| Influenza A virus (A/dove/Korea/S11/03(H3N2))
PB2 (PB2) gene, partial cds. gi|5805276|AF144300| Influenza A virus
(A/Goose/Guangdong/1/96(H5N1)) polymerase (PB2) gene, complete cds.
gi|3335416|AF046086| Influenza A virus (A/Chicken/Hong Kong/220/97
(H5N1)) polymerase basic protein 2 (PB2) gene, partial cds.
gi|6048841|AF098577| Influenza A virus (A/Chicken/Hong Kong/258/97
(H5N1)) PB2 protein (PB2) gene, partial cds. gi|6048843|AF098578|
Influenza A virus (A/Chicken/Hong Kong/y388/97 (H5N1)) PB2 protein
(PB2) gene, partial cds. gi|6048845|AF098579| Influenza A virus
(A/Chicken/Hong Kong/728/97 (H5N1)) PB2 protein (PB2) gene, partial
cds. gi|6048847|AF098580| Influenza A virus (A/Chicken/Hong
Kong/786/97 (H5N1)) PB2 protein (PB2) gene, partial cds.
gi|6048849|AF098581| Influenza A virus (A/Chicken/Hong Kong/915/97
(H5N1)) PB2 protein (PB2) gene, partial cds. gi|6048851|AF098582|
Influenza A virus (A/Duck/Hong Kong/p46/97 (H5N1)) PB2 protein
(PB2) gene, partial cds. gi|6048853|AF098583| Influenza A virus
(A/Duck/Hong Kong/y283/97 (H5N1)) PB2 protein (PB2) gene, partial
cds. gi|6048855|AF098584| Influenza A virus (A/Goose/Hong
Kong/w355/97 (H5N1)) PB2 protein (PB2) gene, partial cds.
gi|14860983|AY038798| Influenza A virus
(A/goose/Guangdong/3/1997(H5N1)) PB2 protein (PB2) gene, complete
cds. gi|47156244|AY585513| Influenza A virus
(A/duck/Guangxi/07/1999(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|9863884|AF216717| Influenza A virus
(A/Environment/Hong Kong/437-4/99 (H5N1)) polymerase basic protein
2 gene, partial cds. gi|9863903|AF216725| Influenza A virus
(A/Environment/Hong Kong/437-6/99 (H5N1)) polymerase basic protein
2 gene, partial cds. gi|9863921|AF216733| Influenza A virus
(A/Environment/Hong Kong/437-8/99 (H5N1)) polymerase basic protein
2 gene, partial cds. gi|9863939|AF216741| Influenza A virus
(A/Environment/Hong Kong/437-10/99 (H5N1)) polymerase basic protein
2 gene, partial cds. gi|47156264|AY585523| Influenza A virus
(A/duck/Zhejiang/11/2000 (H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156266|AY585524| Influenza A virus
(A/duck/Zhejiang/52/2000 (H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156232|AY585507| Influenza A virus
(A/duck/Fujian/19/2000(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156236|AY585509| Influenza A virus
(A/duck/Guangdong/07/2000(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156238|AY585510| Influenza A virus
(A/duck/Guangdong/12/2000(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156242|AY585512| Influenza A virus
(A/duck/Guangdong/40/2000(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|19697849|AY059520| Influenza A virus
(A/Goose/Hong Kong/ww26/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697851|AY059521| Influenza A virus
(A/Goose/Hong Kong/ww28/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697853|AY059522| Influenza A virus
(A/Duck/Hong Kong/ww381/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697855|AY059523| Influenza A virus
(A/Duck/Hong Kong/ww461/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697857|AY059524| Influenza A virus
(A/Goose/Hong Kong/ww491/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697859|AY059525| Influenza A virus
(A/Duck/Hong Kong/2986.1/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|19697867|AY059529| Influenza A virus
(A/Goose/Hong Kong/3014.8/2000(H5N1)) segment 1 polymerase (PB2)
gene, partial cds. gi|18092181|AF398425| Influenza A virus
(A/Goose/Hong Kong/385.3/2000(H5N1)) polymerase (PB2) gene, partial
cds. gi|18092183|AF398426| Influenza A virus (A/Goose/Hong
Kong/385.5/2000(H5N1)) polymerase (PB2) gene, partial cds.
gi|21359665|AF468840| Influenza A virus
(A/duck/Anyang/AVL-1/2001(H5N1)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|28849606|AF509143| Influenza A virus
(A/Chicken/Hong Kong/FY77/01 (H5N1)) polymerase (PB2) gene, partial
cds. gi|28849608|AF509144| Influenza A virus (A/Chicken/Hong
Kong/YU562/01
(H5N1)) polymerase (PB2) gene, partial cds. gi|28849610|AF509145|
Influenza A virus (A/Chicken/Hong Kong/YU563/01 (H5N1)) polymerase
(PB2) gene, partial cds. gi|28849612|AF509146| Influenza A virus
(A/Chicken/Hong Kong/FY150/01 (H5N1)) polymerase (PB2) gene,
partial cds. gi|28849614|AF509147| Influenza A virus
(A/Pheasant/Hong Kong/FY155/01 (H5N1)) polymerase (PB2) gene,
partial cds. gi|28849616|AF509148| Influenza A virus (A/Silky
Chicken/Hong Kong/SF189/01 (H5N1)) polymerase (PB2) gene, partial
cds. gi|28849618|AF509149| Influenza A virus (A/Quail/Hong
Kong/SF203/01 (H5N1) polymerase (PB2) gene, partial cds.
gi|28849620|AF509150| Influenza A virus (A/Pigeon/Hong
Kong/SF215/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849622|AF509151| Influenza A virus (A/Chicken/Hong
Kong/SF219/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849624|AF509152| Influenza A virus (A/Chicken/Hong
Kong/715.5/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849626|AF509153| Influenza A virus (A/Chicken/Hong
Kong/751.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849628|AF509154| Influenza A virus (A/Chicken/Hong
Kong/822.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849630|AF509155| Influenza A virus (A/Chicken/Hong
Kong/829.2/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849632|AF509156| Influenza A virus (A/Chicken/Hong
Kong/830.2/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849634|AF509157| Influenza A virus (A/Chicken/Hong
Kong/858.3/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849636|AF509158| Influenza A virus (A/Chicken/Hong
Kong/866.3/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849638|AF509159| Influenza A virus (A/Chicken/Hong
Kong/867.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849640|AF509160| Influenza A virus (A/Chicken/Hong
Kong/879.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849642|AF509161| Influenza A virus (A/Chicken/Hong
Kong/873.3/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849644|AF509162| Influenza A virus (A/Chicken/Hong
Kong/876.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849646|AF509163| Influenza A virus (A/Chicken/Hong
Kong/891.1/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849648|AF509164| Influenza A virus (A/Chicken/Hong
Kong/893.2/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28849650|AF509165| Influenza A virus (A/Goose/Hong Kong/76.1/01
(H5N1)) polymerase (PB2) gene, partial cds. gi|28849652|AF509166|
Influenza A virus (A/Goose/Hong Kong/ww100/01 (H5N1)) polymerase
(PB2) gene, partial cds. gi|28849654|AF509167| Influenza A virus
(A/Duck/Hong Kong/573.4/01 (H5N1)) polymerase (PB2) gene, partial
cds. gi|28849656|AF509168| Influenza A virus (A/Duck/Hong
Kong/646.3/01 (H5N1)) polymerase (PB2) gene, partial cds.
gi|28823262|AY221584| Influenza A virus
(A/Chicken/HongKong/NT873.3/01- MB(H5N1)) polymerase basic protein
2 (PB2) gene, partial cds. gi|28823443|AY221585| Influenza A virus
(A/Chicken/HongKong/NT873.3/01(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28823612|AY221586| Influenza A virus
(A/Chicken/HongKong/FY150/01- MB(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28823783|AY221587| Influenza A virus
(A/Chicken/HongKong/FY150/01(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28823961|AY221588| Influenza A virus
(A/Pheasant/HongKong/FY155/01- MB(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28824143|AY221589| Influenza A virus
(A/Pheasant/HongKong/FY155/01(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28824334|AY221590| Influenza A virus
(A/Chicken/HongKong/YU822.2/01- MB(H5N1)) polymerase basic protein
2 (PB2) gene, partial cds. gi|28824502|AY221591| Influenza A virus
(A/Chicken/HongKong/YU822.2/01(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|28824684|AY221592| Influenza A virus
(A/Chicken/HongKong/YU562/01(H5N1)) polymerase basic protein 2
(PB2) gene, partial cds. gi|47156230|AY585506| Influenza A virus
(A/duck/Fujian/17/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156234|AY585508| Influenza A virus
(A/duck/Guangdong/01/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156246|AY585514| Influenza A virus
(A/duck/Guangxi/22/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156248|AY585515| Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156250|AY585516| Influenza A virus
(A/duck/Guangxi/50/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156254|AY585518| Influenza A virus
(A/duck/Shanghai/08/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156256|AY585519| Influenza A virus
(A/duck/Shanghai/13/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156262|AY585522| Influenza A virus
(A/duck/Shanghai/38/2001(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156258|AY585520| Influenza A virus
(A/duck/Shanghai/35/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156260|AY585521| Influenza A virus
(A/duck/Shanghai/37/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156252|AY585517| Influenza A virus
(A/duck/Guangxi/53/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156240|AY585511| Influenza A virus
(A/duck/Guangdong/22/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47834791|AY576382| Influenza A virus
(A/Gs/HK/739.2/02 (H5N1)) polymerase (PB2) gene, partial cds.
gi|47834793|AY576383| Influenza A virus (A/Eg/HK/757.3/02 (H5N1))
polymerase (PB2) gene, partial cds. gi|47834795|AY576384| Influenza
A virus (A/G.H/HK/793.1/02 (H5N1)) polymerase (PB2) gene, partial
cds. gi|47834797|AY576385| Influenza A virus (A/Dk/HK/821/02
(H5N1)) polymerase (PB2) gene, partial cds. gi|47834799|AY576386|
Influenza A virus (A/Ck/HK/31.4/02 (H5N1)) polymerase (PB2) gene,
partial cds. gi|47834801|AY576387| Influenza A virus
(A/Ck/HK/61.9/02 (H5N1)) polymerase (PB2) gene, partial cds.
gi|47834803|AY576388| Influenza A virus (A/Ck/HK/YU777/02 (H5N1))
polymerase (PB2) gene, partial cds. gi|47834805|AY576389| Influenza
A virus (A/Ck/HK/96.1/02 (H5N1)) polymerase (PB2) gene, partial
cds. gi|47834807|AY576390| Influenza A virus (A/Ck/HK/409.1/02
(H5N1)) polymerase (PB2) gene, partial cds. gi|47834809|AY576391|
Influenza A virus (A/Ph/HK/sv674.15/02 (H5N1)) polymerase (PB2)
gene, partial cds. gi|47156226|AY585504| Influenza A virus
(A/duck/Fujian/01/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|47156228|AY585505| Influenza A virus
(A/duck/Fujian/13/2002(H5N1)) polymerase basic protein 2 (PB2)
mRNA, complete cds. gi|50296597|AY651744| Influenza A virus (A/grey
heron/HK/861.1/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296599|AY651745| Influenza A virus (A/feral
pigeon/HK/862.7/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296601|AY651746| Influenza A virus (A/tree
sparrow/HK/864/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296603|AY651747| Influenza A virus
(A/teal/China/2978.1/2002(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|56548879|AY676021| Influenza A virus
(A/duck/Hong Kong/821/02(H5N1)) polymerase basic 2 (PB2) gene,
complete cds. gi|50296569|AY651730| Influenza A virus
(A/Gf/HK/38/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296571|AY651731| Influenza A virus
(A/Ck/HK/31.2/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296573|AY651732| Influenza A virus
(A/Ck/HK/37.4/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296575|AY651733| Influenza A virus
(A/SCk/HK/YU100/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296577|AY651734| Influenza A virus
(A/Ck/HK/YU22/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296579|AY651735| Influenza A virus
(A/Ck/HK/3176.3/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296581|AY651736| Influenza A virus
(A/Ck/HK/3169.1/2002(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296583|AY651737| Influenza A virus
(A/Ck/HK/FY157/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296585|AY651738| Influenza A virus
(A/Ck/HK/YU324/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296587|AY651739| Influenza A virus
(A/Ck/HK/2133.1/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296589|AY651740| Influenza A virus
(A/Ck/HK/NT93/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296591|AY651741| Influenza A virus
(A/Ck/HK/SSP141/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296593|AY651742| Influenza A virus
(A/Ck/HK/WF157/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296595|AY651743| Influenza A virus (A/black
headed gull/HK/12.1/2003(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|50296607|AY651749| Influenza A virus
(A/Dk/HN/5806/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296609|AY651750| Influenza A virus
(A/Dk/ST/4003/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296611|AY651751| Influenza A virus
(A/Ck/ST/4231/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296613|AY651752| Influenza A virus
(A/Dk/YN/6255/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296615|AY651753| Influenza A virus
(A/Dk/YN/6445/2003(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296529|AY651710| Influenza A virus
(A/Ck/Indonesia/2A/2003(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|56548881|AY676022| Influenza A virus
(A/egret/Hong Kong/757.2/03(H5N1)) polymerase basic 2 (PB2) gene,
complete cds. gi|56548883|AY676023| Influenza A virus
(A/chicken/Korea/ES/03(H5N1)) polymerase basic 2 (PB2) gene,
complete cds. gi|56548885|AY676024| Influenza A virus
(A/duck/Korea/ESD1/03(H5N1)) polymerase basic 2 (PB2) gene,
complete cds. gi|50296519|AY651705| Influenza A virus
(A/Ck/Indonesia/PA/2003(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296523|AY651707| Influenza A virus
(A/Ck/Indonesia/BL/2003(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|41207501|AY518367| Influenza A virus
(A/duck/China/E319-2/03(H5N1)) polymerase subunit PB2 (PB2) gene,
complete cds. gi|45359369|AY550147| Influenza A virus
(A/chicken/Nakorn-Patom Thailand/CU-K2/04(H5N1)) polymerase basic
protein 2 (PB2) gene, partial cds. gi|50296525|AY651708| Influenza
A virus (A/Ck/Indonesia/5/2004(H5N1)) polymerase basic subunit 2
(PB2) gene, partial cds. gi|50296527|AY651709| Influenza A virus
(A/Ck/Indonesia/4/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|50296521|AY651706| Influenza A virus
(A/Dk/Indonesia/MS/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|51094103|AY590581| Influenza A virus
(A/chicken/Nakorn- Patom/Thailand/CU-K2/2004(H5N1)) polymerase
basic protein 2 (PBP2) gene, partial cds. gi|47716766|AY609309|
Influenza A virus (A/chicken/Guangdong/174/04(H5N1)) segment 1,
complete sequence. gi|58531082|AB166859| Influenza A virus
(A/chicken/Yamaguchi/7/2004(H5N1)) PB2 gene for polymerase basic
protein 2, complete cds. gi|58531114|AB188813| Influenza A virus
(A/chicken/Oita/8/2004(H5N1)) PB2 gene for polymerase basic protein
2, complete cds. gi|50956621|AY684703| Influenza A virus
(A/chicken/Hubei/327/2004(H5N1)) polymerase basic protein 2 (PB2)
gene, complete cds. gi|57915957|AY737286| Influenza A virus
(A/chicken/Guangdong/191/04(H5N1)) segment 1, complete sequence.
gi|57916006|AY737293| Influenza A virus
(A/chicken/Guangdong/178/04(H5N1)) segment 1, complete sequence.
gi|57916060|AY737301| Influenza A virus
(A/duck/Guangdong/173/04(H5N1)) segment 1, complete sequence.
gi|55233237|AY770084| Influenza A virus
(A/chicken/Hubei/489/2004(H5N1)) polymerase basic protein 2 (PB2)
gene, complete cds. gi|54873461|AY770993| Influenza A virus
(A/chicken/Ayutthaya/Thailand/CU- 23/04(H5N1)) polymerase basic
protein 2 gene, partial cds. gi|58618421|AY818127| Influenza A
virus (A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PB2 gene,
complete cds. gi|58618423|AY818128| Influenza A virus
(A/quail/Vietnam/36/04(H5N1)) polymerase protein PB2 gene, complete
cds. gi|58374183|AY856861| Influenza A virus
(A/duck/Shandong/093/2004(H5N1)) segment 1, complete sequence.
gi|58531132|AB188821| Influenza A virus
(A/chicken/Kyoto/3/2004(H5N1)) PB2 gene for polymerase basic
protein 2, complete cds. gi|58531150|AB189050| Influenza A virus
(A/crow/Kyoto/53/2004(H5N1)) PB2 gene for polymerase basic protein
2, complete cds, gi|58531168|AB189058| Influenza A virus
(A/crow/Osaka/102/2004(H5N1)) PB2 gene for polymerase basic protein
2, complete cds, gi|50296605|AY651748| Influenza A virus
(A/peregrine falcon/HK/D0028/2004(H5N1)) polymerase basic subunit 2
(PB2) gene, complete cds.
gi|50296531|AY651711| Influenza A virus
(A/Ck/Thailand/1/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296533|AY651712| Influenza A virus
(A/Ck/Thailand/73/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|50296535|AY651713| Influenza A virus
(A/Ck/Thailand/9.1/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296537|AY651714| Influenza A virus
(A/Qa/Thailand/57/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296539|AY651715| Influenza A virus
(A/bird/Thailand/3.1/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296541|AY651716| Influenza A virus
(A/Dk/Thailand/71.1/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296543|AY651717| Influenza A virus
(A/Gs/Thailand/79/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|50296553|AY651722| Influenza A virus
(A/Ck/Viet Nam/33/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296555|AY651723| Influenza A virus
(A/Ck/Viet Nam/35/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296557|AY651724| Influenza A virus
(A/Ck/Viet Nam/36/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296559|AY651725| Influenza A virus
(A/Ck/Viet Nam/37/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296561|AY651726| Influenza A virus
(A/Ck/Viet Nam/38/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296563|AY651727| Influenza A virus
(A/Ck/Viet Nam/39/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296565|AY651728| Influenza A virus
(A/Ck/Viet Nam/C57/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296567|AY651729| Influenza A virus
(A/Dk/Viet Nam/11/2004(H5N1)) polymerase basic subunit 2 (PB2)
gene, complete cds. gi|50296617|AY651754| Influenza A virus
(A/Ck/YN/374/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296619|AY651755| Influenza A virus
(A/Ck/YN/115/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296621|AY651756| Influenza A virus
(A/Ph/ST/44/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296623|AY651757| Influenza A virus
(A/Dk/HN/303/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50296625|AY651758| Influenza A virus
(A/Dk/HN/101/2004(H5N1)) polymerase basic subunit 2 (PB2) gene,
partial cds. gi|50365712|AY653193| Influenza A virus
(A/chicken/Jilin/9/2004(H5N1)) segment 1, complete sequence.
gi|47680940|AY586445| Influenza A Virus
(A/mallard/Italy/43/01(H7N3)) PB2 gene, partial cds.
gi|47680930|AY586440| Influenza A virus
(A/mallard/Italy/33/01(H7N3)) PB2 gene, partial cds.
gi|47680932|AY586441| Influenza A virus
(A/turkey/Italy/220158/2002(H7N3)) PB2 gene, partial cds.
gi|45124743|AJ627485| Influenza A virus
(A/turkey/Italy/214845/2002(H7N3)) PB2 gene for RNA polymerase,
genomic RNA. gi|45124767|AJ627496| Influenza A virus
(A/turkey/Italy/220158/2002(H7N3)) PB2 gene for RNA polymerase,
genomic RNA. gi|34597782|AY303665| Influenza A virus
(A/chicken/Chile/176822/02(H7N3)) polymerase basic protein 2 gene,
partial cds. gi|34597784|AY303666| Influenza A virus
(A/chicken/Chile/4957/02(H7N3)) polymerase basic protein 2 gene,
partial cds. gi|47680928|AY586439| Influenza A Virus
(A/turkey/Italy/214845/02(H7N3)) PB2 gene, partial cds.
gi|47834374|AY616766| Influenza A virus (A/chicken/British
Columbia/04(H7N3)) PB2 polymerase subunit (PB2) gene, complete cds.
gi|50542651|AY646085| Influenza A virus (A/chicken/British
Columbia/GSC_human_B/04(H7N3)) polymerase basic protein 2 (PB2)
gene, complete cds. gi|50083053|AY648294| Influenza A virus
(A/GSC_chicken_B/British Columbia/04(H7N3)) PB2 polymerase subunit
(PB2) gene, complete cds. gi|50059194|AY650276| Influenza A virus
(A/GSC_chicken/British Columbia/04(H7N3)) PB2 polymerase subunit
(PB2) gene, complete cds. gi|60700|X58691| Influenza A virus
(A/FPV/Dobson/27 (H7N7)) gene for cap- binding protein PB2, genomic
RNA gi|325001|M38291| Influenza virus A/FPV/Weybridge polymerase
basic 2 protein (PB2) (seg 3) gene, complete cds.
gi|9988661|AF268120| Influenza A virus
(A/RedKnot/Delaware/259/94(H7N7)) polymerase protein PB2 gene,
partial cds. gi|40732893|AJ620347| Influenza A virus
((A/Chicken/Germany/R28/03(H7N7)) A/Chicken/Germany/R28/03(H7N7))
PB2 gene for RNA polymerase, genomic RNA. gi|37813157|AY342410|
Influenza A virus (A/Netherlands/124/03(H7N7)) polymerase protein 2
gene, partial cds. gi|37813159|AY342411| Influenza A virus
(A/Netherlands/126/03(H7N7)) polymerase protein 2 gene, partial
cds. gi|37813161|AY342412| Influenza A virus
(A/Netherlands/127/03(H7N7)) polymerase protein 2 gene, partial
cds. gi|37813163|AY342413| Influenza A virus
(A/Netherlands/219/03(H7N7)) polymerase protein 2 gene, partial
cds. gi|37813165|AY342414| Influenza A virus
(A/chicken/Netherlands/1/03(H7N7)) polymerase protein 2 gene,
partial cds. gi|5732354|AF156443| Influenza A virus
(A/Turkey/California/189/66(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|31339587|AF523469| Influenza A virus
(A/Duck/Hong Kong/86/76(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339583|AF523467| Influenza A virus (A/Duck/Hong
Kong/366/78(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339605|AF523478| Influenza A virus (A/Duck/Hong
Kong/289/78(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339585|AF523468| Influenza A virus (A/Duck/Hong
Kong/552/79(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339589|AF523470| Influenza A virus (A/Duck/Hong
Kong/610/79(H9N2)) polymerase (PB2) gene, partial cds.
gi|49356935|AY633283| Influenza A virus
(A/mallard/Alberta/321/88(H9N2)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|49356939|AY633299| Influenza A virus
(A/mallard/Alberta/11/91(H9N2)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|49356907|AY633171| Influenza A virus
(A/mallard/Alberta/17/91(H9N2)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|5732342|AF156437| Influenza A virus
(A/Quail/Hong Kong/AF157/92(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|5732352|AF156442| Influenza A virus
(A/Quail/Arkansas/29209-1/93(H9N2)) segment 1 PB2 polymerase
subunit (PB2) gene, partial cds. gi|5732340|AF156436| Influenza A
virus (A/Chicken/Hong Kong/739/94(H9N2)) segment 1 PB2 polymerase
subunit (PB2) gene, partial cds. gi|5732344|AF156438| Influenza A
virus (A/Chicken/Beijing/1/94(H9N2)) segment 1 PB2 polymerase
subunit (PB2) gene, partial cds. gi|22759060|AF536679| Influenza A
virus (A/Chicken/Beijing/1/95(H9N2)) PB2 gene, partial cds.
gi|33318110|AF508640| Influenza A virus (A/Ostrich/South
Africa/9508103/95(H9N2)) segment 1 polymerase PB2 (PB2) gene,
complete cds. gi|33318118|AF508644| Influenza A virus
(A/Duck/Germany/113/95(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|33318144|AF508657| Influenza A virus
(A/Chicken/Shandong/6/96(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|33318152|AF508661| Influenza A virus
(A/Quail/Shanghai/8/96(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|5732346|AF156439| Influenza A virus
(A/Chicken/Korea/38349- p96323/96(H9N2)) segment 1 PB2 polymerase
subunit (PB2) gene, partial cds. gi|5732348|AF156440| Influenza A
virus (A/Chicken/Korea/25232- 96006/96(H9N2)) segment 1 PB2
polymerase subunit (PB2) gene, partial cds. gi|5732350|AF156441|
Influenza A virus (A/Shorebird/Delaware/9/96(H9N2)) segment 1 PB2
polymerase subunit (PB2) gene, partial cds. gi|22759068|AF536683|
Influenza A virus (A/Chicken/Hebei/1/96(H9N2)) PB2 gene, partial
cds. gi|5732328|AF156430| Influenza A virus (A/Chicken/Hong
Kong/G9/97(H9N2)) segment 1 PB2 polymerase subunit (PB2) gene,
complete cds. gi|5732330|AF156431| Influenza A virus
(A/Chicken/Hong Kong/G23/97(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|5732332|AF156432| Influenza A virus
(A/Pigeon/Hong Kong/Y233/97(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|5732334|AF156433| Influenza A virus
(A/Duck/Hong Kong/Y280/97(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|5732336|AF156434| Influenza A virus
(A/Duck/Hong Kong/Y439/97(H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|5732338|AF156435| Influenza A virus
(A/Quail/Hong Kong/G1/97 (H9N2)) segment 1 PB2 polymerase subunit
(PB2) gene, partial cds. gi|33318148|AF508659| Influenza A virus
(A/Chicken/Shenzhen/9/97(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|33318150|AF508660| Influenza A virus
(A/Duck/Nanjing/1/97(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|33318124|AF508647| Influenza A virus
(A/Pheasant/Ireland/PV18/97(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|13383266|AB049153| Influenza A virus
(A/parakeet/Chiba/1/97(H9N2)) PB2 gene for polymerase basic protein
2, complete cds. gi|33318132|AF508651| Influenza A virus
(A/Chicken/Guangdong/11/97(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|33318136|AF508653| Influenza A virus
(A/Chicken/Heilongjiang/10/97(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|22759062|AF536680| Influenza A virus
(A/Chicken/Beijing/2/97(H9N2)) PB2 gene, partial cds.
gi|33318142|AF508656| Influenza A virus
(A/Chicken/Sichuan/5/97(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|22759066|AF536682| Influenza A virus
(A/Chicken/Guangdong/97(H9N2)) PB2 gene, partial cds.
gi|22759078|AF536688| Influenza A virus
(A/Chicken/Shandong/98(H9N2)) PB2 gene, partial cds.
gi|33318116|AF508643| Influenza A virus
(A/Chicken/Germany/R45/98(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|33318134|AF508652| Influenza A virus
(A/Chicken/Hebei/4/98(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|33318128|AF508649| Influenza A virus
(A/Chicken/Beijing/8/98(H9N2)) segment 1 polymerase PB2 (PB2) gene,
complete cds. gi|13383268|AB049154| Influenza A virus
(A/parakeet/Narita/92A/98(H9N2)) PB2 gene for polymerase basic
protein 2, complete cds. gi|22759070|AF536684| Influenza A virus
(A/Chicken/Hebei/2/98(H9N2)) PB2 gene, partial cds.
gi|22759072|AF536685| Influenza A virus
(A/Chicken/Hebei/3/98(H9N2)) PB2 gene, partial cds.
gi|22759074|AF536686| Influenza A virus (A/Chicken/Henan/98(H9N2))
PB2 gene, partial cds. gi|30025722|AY253750| Influenza A virus
(A/Chicken/Shanghai/F/98(H9N2)) RNA polymerase (PB2) gene, complete
cds. gi|12060631|AF222622| Influenza A virus (A/Quail/Hong
Kong/A17/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060633|AF222623| Influenza A virus (A/Pigeon/Hong
Kong/FY6/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060635|AF222624| Influenza A virus (A/Chicken/Hong
Kong/NT16/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060637|AF222625| Influenza A virus (A/Quail/Hong
Kong/SSP10/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial
cds. gi|12060639|AF222626| Influenza A virus (A/Pheasant/Hong
Kong/SSP11/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial
cds. gi|12060641|AF222627| Influenza A virus (A/Chicken/Hong
Kong/FY20/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060643|AF222628| Influenza A virus (A/Chicken/Hong
Kong/KC12/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060645|AF222629| Influenza A virus (A/Quail/Hong
Kong/NT28/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060647|AF222630| Influenza A virus (A/Chicken/Hong
Kong/SF2/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|12060649|AF222631| Influenza A virus (A/Silky Chicken/Hong
Kong/SF44/99(H9N2)) segment 1 polymerase 2 (PB2) gene, partial cds.
gi|22759076|AF536687| Influenza A virus
(A/Chicken/Liaoning/99(H9N2)) PB2 gene, partial cds.
gi|33318112|AF508641| Influenza A virus
(A/Chicken/Pakistan/4/99(H9N2)) segment 1 polymerase PB2 (PB2)
gene, complete cds. gi|33318114|AF508642| Influenza A virus
(A/Chicken/Pakistan/5/99(H9N2)) segment 1 polymerase PB2 (PB2)
gene, complete cds. gi|33318126|AF508648| Influenza A virus
(A/Chicken/Korea/99029/99(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|33318120|AF508645| Influenza A virus
(A/Chicken/Iran/11T/99(H9N2)) segment 1 polymerase PB2 (PB2) gene,
complete cds. gi|33318122|AF508646| Influenza A virus
(A/Chicken/Saudi Arabia/532/99(H9N2)) segment 1 polymerase PB2
(PB2) gene, partial cds. gi|33318140|AF508655| Influenza A virus
(A/Chicken/Ningxia/5/99(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|33318146|AF508658| Influenza A virus
(A/Chicken/Shijiazhuang/2/99(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|12038893|AJ291395| Influenza A virus
(A/Chicken/Pakistan/2/99(H9N2)) PB2 gene for polymerase PB2,
genomic RNA. gi|22759064|AF536681| Influenza A virus
(A/Chicken/Beijing/3/99(H9N2)) PB2 gene, partial cds.
gi|31339607|AF523479| Influenza A virus
(A/Duck/Shantou/1881/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339601|AF523476| Influenza A virus
(A/Duck/Shantou/830/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339603|AF523477| Influenza A virus
(A/Duck/Shantou/1796/00(H9N2))
polymerase (PB2) gene, partial cds. gi|18496117|AJ427861| Influenza
A virus (A/quail/Hong Kong/FY298/00(H9N2)) partial pb2 gene for PB2
polymerase protein, genomic RNA gi|27466041|AY180715| Influenza A
virus strain A/Wild Duck/Nanchang/2- 0480/2000 (H9N2) polymerase
subunit PB2 (PB2) gene, partial cds. gi|27466043|AY180716|
Influenza A virus strain A/Pigeon/Nanchang/2- 0461/2000 (H9N2)
polymerase subunit PB2 (PB2) gene, partial cds.
gi|27466045|AY180717| Influenza A virus strain
A/Duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466055|AY180722| Influenza A virus strain
A/Duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466057|AY180723| Influenza A virus strain
A/Pigeon/Nanchang/7-058/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466067|AY180728| Influenza A virus strain
A/Quail/Nanchang/2-0460/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466085|AY180737| Influenza A virus strain
A/Pigeon/Nanchang/11- 145/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466087|AY180738| Influenza A virus strain
A/Duck/Nanchang/11-197/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466091|AY180740| Influenza A virus strain
A/Duck/Nanchang/11-290/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466093|AY180741| Influenza A virus strain
A/Duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|31339575|AF523463| Influenza A virus
(A/Duck/Shantou/2134/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339579|AF523465| Influenza A virus
(A/Duck/Shantou/1043/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339581|AF523466| Influenza A virus
(A/Duck/Shantou/1042/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339593|AF523472| Influenza A virus
(A/Duck/Shantou/2102/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339595|AF523473| Influenza A virus
(A/Duck/Shantou/2144/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339597|AF523474| Influenza A virus
(A/Duck/Shantou/2143/00(H9N2)) polymerase (PB2) gene, partial cds.
gi|33318138|AF508654| Influenza A virus
(A/Chicken/Henan/62/00(H9N2)) segment 1 polymerase PB2 (PB2) gene,
partial cds. gi|33318130|AF508650| Influenza A virus
(A/Chicken/Guangdong/10/00(H9N2)) segment 1 polymerase PB2 (PB2)
gene, partial cds. gi|27466121|AY180755| Influenza A virus strain
A/Duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466141|AY180765| Influenza A virus strain
A/Chicken/Nanchang/4- 010/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466157|AY180773| Influenza A virus strain
A/Quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|27466159|AY180774| Influenza A virus strain
A/Chicken/Nanchang/1- 0016/2000 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|55469788|AY768575| Influenza A virus
(A/chicken/Korea/SNU0028/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469790|AY768576| Influenza A virus
(A/chicken/Korea/SNU0037/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469792|AY768577| Influenza A virus
(A/chicken/Korea/SNU0073/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469794|AY768578| Influenza A virus
(A/chicken/Korea/SNU0091/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469796|AY768579| Influenza A virus
(A/chicken/Korea/SNU0140/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469798|AY768580| Influenza A virus
(A/chicken/Korea/SNU0146/00(H9N2)) polymerase basic subunit 2 (PB2)
gene, partial cds. gi|55469800|AY768581| Influenza A virus
(A/chicken/Korea/SNU1035C/00(H9N2)) polymerase basic subunit 2
(PB2) gene, partial cds. gi|27466143|AY180766| Influenza A virus
strain A/Chicken/Nanchang/4- 301/2001 (H9N2) polymerase subunit PB2
(PB2) gene, partial cds. gi|31339599|AF523475| Influenza A virus
(A/Duck/Shantou/2088/01(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339591|AF523471| Influenza A virus
(A/Duck/Shantou/1605/01(H9N2)) polymerase (PB2) gene, partial cds.
gi|31339577|AF523464| Influenza A virus (A/Wild
Duck/Shantou/4808/01(H9N2)) polymerase (PB2) gene, partial cds.
gi|27466097|AY180743| Influenza A virus strain
A/Chicken/Nanchang/4- 361/2001 (H9N2) polymerase subunit PB2 (PB2)
gene, partial cds. gi|54398631|AY664792| Influenza A virus
(A/chicken/HongKong/CSW153/03(H9N2)) polymerase basic protein 2
(PB2) gene, partial cds. gi|54398633|AY664793| Influenza A virus
(A/chicken/HongKong/AP45/03(H9N2)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|54398635|AY664794| Influenza A virus
(A/chicken/HongKong/BD90/03(H9N2)) polymerase basic protein 2 (PB2)
gene, partial cds. gi|54398637|AY664795| Influenza A virus
(A/chicken/HongKong/CSW291/03(H9N2)) nonfunctional polymerase basic
protein 2 (PB2) gene, partial sequence. gi|54398638|AY664796|
Influenza A virus (A/chicken/HongKong/CSW304/03(H9N2))
nonfunctional polymerase basic protein 2 (PB2) gene, partial
sequence. gi|54398639|AY664797| Influenza A virus
(A/chicken/HongKong/FY23/03(H9N2)) nonfunctional polymerase basic
protein 2 (PB2) gene, partial sequence. gi|54398640|AY664798|
Influenza A virus (A/guineafowl/HongKong/NT101/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|54398642|AY664799|
Influenza A virus (A/chicken/HongKong/NT142/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|54398644|AY664800|
Influenza A virus (A/chicken/HongKong/SF1/03(H9N2)) nonfunctional
polymerase basic protein 2 (PB2) gene, partial sequence.
gi|54398645|AY664801| Influenza A virus
(A/chicken/HongKong/SSP101/03(H9N2)) nonfunctional polymerase basic
protein 2 (PB2) gene, partial sequence. gi|54398646|AY664802|
Influenza A virus (A/chicken/HongKong/TP38/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|54398648|AY664803|
Influenza A virus (A/chicken/HongKong/WF126/03(H9N2)) nonfunctional
polymerase basic protein 2 (PB2) gene, partial sequence.
gi|54398649|AY664804| Influenza A virus
(A/pigeon/HongKong/WF53/03(H9N2)) nonfunctional polymerase basic
protein 2 (PB2) gene, partial sequence. gi|54398650|AY664805|
Influenza A virus (A/pheasant/HongKong/WF54/03(H9N2)) nonfunctional
polymerase basic protein 2 (PB2) gene, partial sequence.
gi|54398651|AY664806| Influenza A virus
(A/guineafowl/HongKong/NT184/03(H9N2)) polymerase basic protein 2
(PB2) gene, partial cds. gi|54398653|AY664807| Influenza A virus
(A/chicken/HongKong/WF120/03(H9N2)) nonfunctional polymerase basic
protein 2 (PB2) gene, partial sequence. gi|54398654|AY664808|
Influenza A virus (A/chicken/HongKong/NT366/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|54398656|AY664809|
Influenza A virus (A/chicken/HongKong/SSP418/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|54398658|AY664810|
Influenza A virus (A/chicken/HongKong/YU427/03(H9N2)) polymerase
basic protein 2 (PB2) gene, partial cds. gi|55793686|AY800240|
Influenza A virus (A/chicken/Korea/S1/2003(H9N2)) polymerase basic
protein 2 (PB2) gene, partial cds. gi|58429686|AY862710| Influenza
A virus (A/silky chicken/Korea/S3/03(H9N2)) PB2 (PB2) gene, partial
cds. gi|58429688|AY862711| Influenza A virus
(A/chicken/Korea/S4/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429690|AY862712| Influenza A virus
(A/chicken/Korea/S5/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429692|AY862713| Influenza A virus
(A/chicken/Korea/S12/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429694|AY862714| Influenza A virus (A/duck/Korea/S13/03(H9N2))
PB2 (PB2) gene, partial cds. gi|58429696|AY862715| Influenza A
virus (A/dove/Korea/S14/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429698|AY862716| Influenza A virus
(A/chicken/Korea/S15/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429700|AY862717| Influenza A virus
(A/chicken/Korea/S16/03(H9N2)) PB2 (PB2) gene, partial cds.
gi|58429702|AY862718| Influenza A virus
(A/chicken/Korea/S18/03(H9N2)) PB2 (PB2) gene, partial cds.
Sequences used in analysis of Influenza A Polymerase Acidic protein
(PA) gi|27465935|AY180662| Influenza A virus strain
A/Quail/Nanchang/12-340/2000 (H1N1) polymerase subunit PA (PA)
gene, partial cds. gi|49357063|AY633217| Influenza A virus
(A/mallard/Alberta/211/98(H1N1)) polymerase protein A (PA) gene,
partial cds. gi|5918195|AJ243994| Influenza A virus (STRAIN
A/MALLARD/NEW YORK/6750/78) partial mRNA for PA protein.
gi|45272157|AY422034| Influenza A virus
(A/Duck/Hokkaido/95/01(H2N2)) PA protein (PA) gene, partial cds.
gi|27465965|AY180677| Influenza A virus strain
A/Chicken/Nanchang/3- 120/2001 (H3N2) polymerase subunit PA (PA)
gene, partial cds. gi|56159994|AY779263| Influenza A virus
(A/turkey/North Carolina/12344/03(H3N2)) polymerase acidic protein
(PA) gene, partial cds. gi|56159996|AY779264| Influenza A virus
(A/turkey/Minnesota/764-2/03(H3N2)) polymerase acidic protein (PA)
gene, partial cds. gi|58429736|AY862687| Influenza A virus
(A/chicken/Korea/S6/03(H3N2)) PA (PA) gene, partial cds.
gi|58429738|AY862688| Influenza A virus (A/duck/Korea/S7/03(H3N2))
PA (PA) gene, partial cds. gi|58429740|AY862689| Influenza A virus
(A/duck/Korea/S8/03(H3N2)) PA (PA) gene, partial cds.
gi|58429742|AY862690| Influenza A virus (A/duck/Korea/S9/03(H3N2))
PA (PA) gene, partial cds. gi|58429744|AY862691| Influenza A virus
(A/duck/Korea/S10/03(H3N2)) PA (PA) gene, partial cds.
gi|58429746|AY862692| Influenza A virus (A/dove/Korea/S11/03(H3N2))
PA (PA) gene, partial cds. gi|18091833|AF213914| Influenza A virus
(A/Chicken/Italy/5945/95(H3N2)) segment 3 PA polymerase protein
gene, partial cds. gi|58531086|AB166861| Influenza A virus
(A/chicken/Yamaguchi/7/2004(H5N1)) PA gene for polymerase acidic
protein, complete cds. gi|58531118|AB188815| Influenza A virus
(A/chicken/Oita/8/2004(H5N1)) PA gene for polymerase acidic
protein, complete cds. gi|9863935|AF216739| Influenza A virus
(A/Environment/Hong Kong/437-10/99 (H5N1)) polymerase acidic
protein gene, complete cds. gi|14165201|AF380163| Influenza A virus
(A/Goose/Guangdong/3/97(H5N1)) segment 3 polymerase (PA) gene,
complete cds. gi|18092185|AF398427| Influenza A virus (A/Goose/Hong
Kong/385.3/2000(H5N1)) polymerase (PA) gene, partial cds.
gi|18092187|AF398428| Influenza A virus (A/Goose/Hong
Kong/385.5/2000(H5N1)) polymerase (PA) gene, partial cds.
gi|21359667|AF468841| Influenza A virus
(A/Duck/Anyang/AVL-1/2001(H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|28849710|AF509195| Influenza A virus
(A/Chicken/Hong Kong/FY77/01 (H5N1)) polymerase (PA) gene, partial
cds. gi|28849712|AF509196| Influenza A virus (A/Chicken/Hong
Kong/YU562/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849714|AF509197| Influenza A virus (A/Chicken/Hong
Kong/YU563/01 (H5N1)) polymerase (PA) gene, complete cds.
gi|28849716|AF509198| Influenza A virus (A/Chicken/Hong
Kong/FY150/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849718|AF509199| Influenza A virus (A/Pheasant/Hong
Kong/FY155/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849720|AF509200| Influenza A virus (A/Silky Chicken/Hong
Kong/SF189/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849722|AF509201| Influenza A virus (A/Quai1/Hong Kong/SF203/01
(H5N1)) polymerase (PA) gene, partial cds. gi|28849724|AF509202|
Influenza A virus (A/Pigeon/Hong Kong/SF215/01 (H5N1)) polymerase
(PA) gene, partial cds. gi|28849726|AF509203| Influenza A virus
(A/Chicken/Hong Kong/SF219/01 (H5N1)) polymerase (PA) gene, partial
cds. gi|28849728|AF509204| Influenza A virus (A/Chicken/Hong
Kong/715.5/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849730|AF509205| Influenza A virus (A/Chicken/Hong
Kong/751.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849732|AF509206| Influenza A virus (A/Chicken/Hong
Kong/822.1/01
(H5N1)) polymerase (PA) gene, partial cds. gi|28849734|AF509207|
Influenza A virus (A/Chicken/Hong Kong/829.2/01 (H5N1)) polymerase
(PA) gene, partial cds. gi|28849736|AF509208| Influenza A virus
(A/Chicken/Hong Kong/830.2/01 (H5N1)) polymerase (PA) gene, partial
cds. gi|28849738|AF509209| Influenza A virus (A/Chicken/Hong
Kong/858.3/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849740|AF509210| Influenza A virus (A/Chicken/Hong
Kong/866.3/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849742|AF509211| Influenza A virus (A/Chicken/Hong
Kong/867.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849744|AF509212| Influenza A virus (A/Chicken/Hong
Kong/879.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849746|AF509213| Influenza A virus (A/Chicken/Hong
Kong/873.3/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849748|AF509214| Influenza A virus (A/Chicken/Hong
Kong/876.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849750|AF509215| Influenza A virus (A/Chicken/Hong
Kong/891.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849752|AF509216| Influenza A virus (A/Chicken/Hong
Kong/893.1/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|28849754|AF509217| Influenza A virus (A/Goose/Hong Kong/76.1/01
(H5N1)) polymerase (PA) gene, partial cds. gi|28849756|AF509218|
Influenza A virus (A/Goose/Hong Kong/ww100/01 (H5N1)) polymerase
(PA) gene, partial cds. gi|28849758|AF509219| Influenza A virus
(A/Duck/Hong Kong/573.4/01 (H5N1)) polymerase (PA) gene, partial
cds. gi|28849760|AF509220| Influenza A virus (A/Duck/Hong
Kong/646.3/01 (H5N1)) polymerase (PA) gene, partial cds.
gi|19697861|AY059526| Influenza A virus (A/Goose/Hong
Kong/ww26/2000(H5N1)) segment 3 polymerase (PA) gene, partial cds.
gi|19697863|AY059527| Influenza A virus (A/Goose/Hong
Kong/ww28/2000(H5N1)) segment 3 polymerase (PA) gene, partial cds.
gi|19697865|AY059528| Influenza A virus (A/Duck/Hong
Kong/ww381/2000(H5N1)) segment 3 polymerase (PA) gene, partial cds.
gi|19697869|AY059530| Influenza A virus (A/Duck/Hong
Kong/ww461/2000(H5N1)) segment 3 polymerase (PA) gene, partial cds.
gi|19697871|AY059531| Influenza A virus (A/Goose/Hong
Kong/ww491/2000(H5N1)) segment 3 polymerase (PA) gene, partial cds.
gi|19697873|AY059532| Influenza A virus (A/Duck/Hong
Kong/2986.1/2000(H5N1)) segment 3 polymerase (PA) gene, partial
cds. gi|19697875|AY059533| Influenza A virus (A/Goose/Hong
Kong/3014.8/2000(H5N1)) segment 3 polymerase (PA) gene, partial
cds. gi|28821204|AY221566| Influenza A virus
(A/Chicken/HongKong/NT873.3/01- MB(H5N1)) polymerase (PA) gene,
partial cds. gi|28821206|AY221567| Influenza A virus
(A/Chicken/HongKong/NT873.3/01(H5N1)) polymerase (PA) gene, partial
cds. gi|28821208|AY221568| Influenza A virus
(A/Chicken/HongKong/FY150/01- MB(H5N1)) polymerase (PA) gene,
partial cds. gi|28821210|AY221569| Influenza A virus
(A/Chicken/HongKong/FY150/01(H5N1)) polymerase (PA) gene, partial
cds. gi|28821212|AY221570| Influenza A virus
(A/Pheasant/HongKong/FY155/01- MB(H5N1)) polymerase (PA) gene,
partial cds. gi|28821214|AY221571| Influenza A virus
(A/Pheasant/HongKong/FY155/01(H5N1)) polymerase (PA) gene, partial
cds. gi|28821216|AY221572| Influenza A virus
(A/Chicken/HongKong/YU822.2/01- MB(H5N1)) polymerase (PA) gene,
partial cds. gi|28821218|AY221573| Influenza A virus
(A/Chicken/HongKong/YU822.2/01(H5N1)) polymerase (PA) gene, partial
cds. gi|28821220|AY221574| Influenza A virus
(A/Chicken/HongKong/YU562/01(H5N1)) polymerase (PA) gene, partial
cds. gi|41207483|AY518365| Influenza A virus
(A/duck/China/E319-2/03(H5N1)) polymerase (PA) gene, complete cds.
gi|51094114|AY551934| Influenza A virus (A/chicken/Nakorn-Patom
Thailand/CU-K2/04(H5N1)) polymerase (PA) gene, complete cds.
gi|47834839|AY576406| Influenza A virus (A/Gs/HK/739.2/02 (H5N1))
polymerase (PA) gene, partial cds. gi|47834841|AY576407| Influenza
A virus (A/Eg/HK/757.3/02 (H5N1)) polymerase (PA) gene, partial
cds. gi|47834843|AY576408| Influenza A virus (A/G.H/HK/793.1/02
(H5N1)) polymerase (PA) gene, partial cds. gi|47834845|AY576409|
Influenza A virus (A/Dk/HK/821/02 (H5N1)) polymerase (PA) gene,
partial cds. gi|47834847|AY576410| Influenza A virus
(A/Ck/HK/31.4/02 (H5N1)) polymerase (PA) gene, complete cds.
gi|47834849|AY576411| Influenza A virus (A/Ck/HK/61.9/02 (H5N1))
polymerase (PA) gene, complete cds. gi|47834851|AY576412| Influenza
A virus (A/Ck/HK/YU777/02 (H5N1)) polymerase (PA) gene, complete
cds. gi|47834853|AY576413| Influenza A virus (A/Ck/HK/96.1/02
(H5N1)) polymerase (PA) gene, partial cds. gi|47834855|AY576414|
Influenza A virus (A/Ck/HK/409.1/02 (H5N1)) polymerase (PA) gene,
partial cds. gi|47834857|AY576415| Influenza A virus
(A/Ph/HK/sv674.15/02 (H5N1)) polymerase (PA) gene, complete cds.
gi|47156478|AY585462| Influenza A virus
(A/duck/Fujian/01/2002(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156480|AY585463| Influenza A virus
(A/duck/Fujian/13/2002(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156482|AY585464| Influenza A virus
(A/duck/Fujian/17/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156484|AY585465| Influenza A virus
(A/duck/Fujian/19/2000(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156486|AY585466| Influenza A virus
(A/duck/Guangdong/01/2001(H5N1)) polymerase (PA) mRNA, complete
cds. gi|47156488|AY585467| Influenza A virus
(A/duck/Guangdong/07/2000(H5N1)) polymerase (PA) mRNA, complete
cds. gi|47156490|AY585468| Influenza A virus
(A/duck/Guangdong/12/2000(H5N1)) polymerase (PA) mRNA, complete
cds. gi|47156492|AY585469| Influenza A virus
(A/duck/Guangdong/22/2002(H5N1)) polymerase (PA) mRNA, complete
cds. gi|47156494|AY585470| Influenza A virus
(A/duck/Guangdong/40/2000(H5N1)) polymerase (PA) mRNA, complete
cds. gi|47156496|AY585471| Influenza A virus
(A/duck/Guangxi/07/1999(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156498|AY585472| Influenza A virus
(A/duck/Guangxi/22/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156500|AY585473| Influenza A virus
(A/duck/Guangxi/35/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156502|AY585474| Influenza A virus
(A/duck/Guangxi/50/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156504|AY585475| Influenza A virus
(A/duck/Guangxi/53/2002(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156506|AY585476| Influenza A virus
(A/duck/Shanghai/08/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156508|AY585477| Influenza A virus
(A/duck/Shanghai/13/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156510|AY585478| Influenza A virus
(A/duck/Shanghai/35/2002(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156512|AY585479| Influenza A virus
(A/duck/Shanghai/37/2002(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156514|AY585480| Influenza A virus
(A/duck/Shanghai/38/2001(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156516|AY585481| Influenza A virus
(A/duck/Zhejiang/11/2000(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47156518|AY585482| Influenza A virus
(A/duck/Zhejiang/52/2000(H5N1)) polymerase (PA) mRNA, complete cds.
gi|47716770|AY609311| Influenza A virus
(A/chicken/Guangdong/174/04(H5N1)) segment 3, complete sequence.
gi|50313026|AY651597| Influenza A virus
(A/Ck/Indonesia/4/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313028|AY651598| Influenza A virus
(A/Ck/Indonesia/5/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313030|AY651599| Influenza A virus
(A/Ck/Indonesia/2A/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313032|AY651600| Influenza A virus
(A/Dk/Indonesia/MS/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313034|AY651601| Influenza A virus
(A/Ck/Indonesia/BL/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313036|AY651602| Influenza A virus
(A/Ck/Indonesia/PA/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313038|AY651603| Influenza A virus
(A/Ck/Thailand/1/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313040|AY651604| Influenza A virus
(A/Ck/Thailand/73/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313042|AY651605| Influenza A virus
(A/Ck/Thailand/9.1/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313044|AY651606| Influenza A virus
(A/Qa/Thailand/57/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313046|AY651607| Influenza A virus
(A/bird/Thailand/3.1/2004(H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|50313048|AY651608| Influenza A virus
(A/Dk/Thailand/71.1/2004(H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|50313050|AY651609| Influenza A virus
(A/Gs/Thailand/79/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313060|AY651614| Influenza A virus (A/Ck/Viet
Nam/33/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313062|AY651615| Influenza A virus (A/Ck/Viet
Nam/35/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313064|AY651616| Influenza A virus (A/Ck/Viet
Nam/36/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313066|AY651617| Influenza A virus (A/Ck/Viet
Nam/37/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313068|AY651618| Influenza A virus (A/Ck/Viet
Nam/38/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313070|AY651619| Influenza A virus (A/Ck/Viet
Nam/39/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313072|AY651620| Influenza A virus (A/Ck/Viet
Nam/C57/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313074|AY651621| Influenza A virus (A/Dk/Viet
Nam/11/2004(H5N1)) polymerase acidic protein (PA) gene, partial
cds. gi|50313076|AY651622| Influenza A virus
(A/Gf/HK/38/2002(H5N1)) polymerase acidic protein (PA) gene,
complete cds. gi|50313078|AY651623| Influenza A virus
(A/Ck/HK/31.2/2002(H5N1)) polymerase acidic protein (PA) gene,
complete cds. gi|50313080|AY651624| Influenza A virus
(A/Ck/HK/37.4/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313082|AY651625| Influenza A virus
(A/SCk/HK/YU100/2002(H5N1)) polymerase acidic protein (PA) gene,
complete cds. gi|50313084|AY651626| Influenza A virus
(A/Ck/HK/YU22/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313086|AY651627| Influenza A virus
(A/Ck/HK/3176.3/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313088|AY651628| Influenza A virus
(A/Ck/HK/3169.1/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313090|AY651629| Influenza A virus
(A/Ck/HK/FY157/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313092|AY651630| Influenza A virus
(A/Ck/HK/YU324/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313094|AY651631| Influenza A virus
(A/Ck/HK/2133.1/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313096|AY651632| Influenza A virus
(A/Ck/HK/NT93/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313098|AY651633| Influenza A virus
(A/Ck/HK/SSP141/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313100|AY651634| Influenza A virus
(A/Ck/HK/WF157/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313102|AY651635| Influenza A virus (A/black
headed gull/HK/12.1/2003(H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|50313104|AY651636| Influenza A virus (A/grey
heron/HK/861.1/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313106|AY651637| Influenza A virus (A/feral
pigeon/HK/862.7/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313108|AY651638| Influenza A virus (A/tree
sparrow/HK/864/2002(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313110|AY651639| Influenza A virus
(A/teal/China/2978.1/2002(H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|50313112|AY651640| Influenza A virus
(A/peregrine falcon/HK/D0028/2004(H5N1)) polymerase acidic protein
(PA) gene, partial cds. gi|50313114|AY651641| Influenza A virus
(A/Dk/HN/5806/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313116|AY651642| Influenza A virus
(A/Dk/HN/303/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313118|AY651643| Influenza A virus
(A/Dk/HN/101/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313120|AY651644| Influenza A virus
(A/Dk/ST/4003/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313122|AY651645| Influenza A virus
(A/Ph/ST/44/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313124|AY651646| Influenza A virus
(A/Ck/ST/4231/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313126|AY651647| Influenza A virus
(A/Dk/YN/6255/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313128|AY651648| Influenza A virus
(A/Dk/YN/6445/2003(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|50313130|AY651649| Influenza A virus
(A/Ck/YN/115/2004(H5N1)) polymerase
acidic protein (PA) gene, partial cds. gi|50313132|AY651650|
Influenza A virus (A/Ck/YN/374/2004(H5N1)) polymerase acidic
protein (PA) gene, partial cds. gi|50365724|AY653198| Influenza A
virus (A/chicken/Jilin/9/2004(H5N1)) segment 3, complete sequence.
gi|56548923|AY676029| Influenza A virus (A/duck/Hong
Kong/821/02(H5N1)) polymerase (PA) gene, complete cds.
gi|56548925|AY676030| Influenza A virus (A/egret/Hong
Kong/757.2/03(H5N1)) polymerase (PA) gene, complete cds.
gi|56548927|AY676031| Influenza A virus
(A/chicken/Korea/ES/03(H5N1)) polymerase (PA) gene, complete cds.
gi|56548929|AY676032| Influenza A virus
(A/duck/Korea/ESD1/03(H5N1)) polymerase (PA) gene, complete cds.
gi|50956625|AY684705| Influenza A virus
(A/chicken/Hubei/327/2004(H5N1)) polymerase (PA) gene, complete
cds. gi|56119221|AY720944| Influenza A virus (A/chicken/Viet
Nam/DT- 171/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|56119227|AY720947| Influenza A virus (A/duck/Viet
Nam/TG- 007A/2004(H5N1)) polymerase acidic protein (PA) gene,
partial cds. gi|57924419|AY724784| Influenza A virus
(A/chicken/Viet Nam/HCM- 022/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57924480|AY724786| Influenza A virus
(A/chicken/Viet Nam/DN- 045/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57924569|AY724788| Influenza A virus
(A/chicken/Viet Nam/VL- 008/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57924680|AY724790| Influenza A virus
(A/chicken/Viet Nam/AG- 010/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57924765|AY724792| Influenza A virus
(A/chicken/Viet Nam/DT- 015/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57924882|AY724796| Influenza A virus
(A/chicken/Viet Nam/LA- 024/2004(H5N1)) polymerase (PA) gene,
partial cds. gi|57915971|AY737288| Influenza A virus
(A/chicken/Guangdong/191/04(H5N1)) segment 3, complete sequence.
gi|57916018|AY737295| Influenza A virus
(A/chicken/Guangdong/178/04(H5N1)) segment 3, complete sequence.
gi|57916074|AY737303| Influenza A virus
(A/duck/Guangdong/173/04(H5N1)) segment 3, complete sequence.
gi|55233234|AY770082| Influenza A virus
(A/chicken/Hubei/489/2004(H5N1)) polymerase (PA) gene, complete
cds. gi|54873465|AY770995| Influenza A virus
(A/chicken/Ayutthaya/Thailand/CU- 23/04(H5N1)) polymerase gene,
partial cds. gi|58618433|AY818133| Influenza A virus
(A/chicken/Vietnam/C58/04(H5N1)) polymerase protein PA gene,
complete cds. gi|58618435|AY818134| Influenza A virus
(A/quail/Vietnam/36/04(H5N1)) polymerase protein PA gene, complete
cds. gi|58374187|AY856863| Influenza A virus
(A/duck/Shandong/093/2004(H5N1)) segment 3, complete sequence.
gi|58531136|AB188823| Influenza A virus
(A/chicken/Kyoto/3/2004(H5N1)) PA gene for polymerase acidic
protein, complete cds. gi|58531154|AB189052| Influenza A virus
(A/crow/Kyoto/53/2004(H5N1)) PA gene for polymerase acidic protein,
complete cds, gi|58531170|AB189059| Influenza A virus
(A/crow/Osaka/102/2004(H5N1)) PA gene for polymerase acidic
protein, complete cds,. gi|3335418|AF046087| Influenza A virus
(A/Chicken/Hong Kong/220/97 (H5N1)) polymerase acidic protein (PA)
gene, partial cds. gi|6048895|AF098604| Influenza A virus
(A/Chicken/Hong Kong/258/97 (H5N1)) PA protein (PA) gene, complete
cds. gi|6048897|AF098605| Influenza A virus (A/Chicken/Hong
Kong/y388/97 (H5N1)) PA protein (PA) gene, complete cds.
gi|6048899|AF098606| Influenza A virus (A/Chicken/Hong Kong/728/97
(H5N1)) PA protein (PA) gene, complete cds. gi|6048901|AF098607|
Influenza A virus (A/Chicken/Hong Kong/786/97 (H5N1)) PA protein
(PA) gene, complete cds. gi|6048903|AF098608| Influenza A virus
(A/Chicken/Hong Kong/915/97 (H5N1)) PA protein (PA) gene, complete
cds. gi|6048905|AF098609| Influenza A virus (A/Duck/Hong
Kong/p46/97 (H5N1)) PA protein (PA) gene, complete cds.
gi|6048907|AF098610| Influenza A virus (A/Duck/Hong Kong/y283/97
(H5N1)) PA protein (PA) gene, complete cds. gi|6048909|AF098611|
Influenza A virus (A/Goose/Hong Kong/w355/97 (H5N1)) PA protein
(PA) gene, complete cds. gi|5805280|AF144302| Influenza A virus
(A/Goose/Guangdong/1/96(H5N1)) polymerase (PA) gene, complete cds.
gi|9863880|AF216715| Influenza A virus (A/Environment/Hong
Kong/437-4/99 (H5N1)) polymerase acidic protein gene, complete cds.
gi|9863899|AF216723| Influenza A virus (A/Environment/Hong
Kong/437-6/99 (H5N1)) polymerase acidic protein gene, complete cds.
gi|9863917|AF216731| Influenza A virus (A/Environment/Hong
Kong/437-8/99 (H5N1)) polymerase acidic protein gene, complete cds.
gi|34597776|AY303660| Influenza A virus
(A/chicken/Chile/176822/02(H7N3)) polymerase acidic protein gene,
complete cds. gi|34597778|AY303661| Influenza A virus
(A/chicken/Chile/4957/02(H7N3)) polymerase acidic protein gene,
complete cds. gi|34597780|AY303662| Influenza A virus
(A/chicken/Chile/4322/02(H7N3)) polymerase acidic protein gene,
partial cds. gi|47680912|AY586431| Influenza A Virus
(A/mallard/Italy/43/01(H7N3)) PA gene, partial cds.
gi|47680914|AY586432| Influenza A virus
(A/mallard/Italy/33/01(H7N3)) PA gene, partial cds.
gi|47680916|AY586433| Influenza A virus
(A/turkey/Italy/220158/2002(H7N3)) PA gene, partial cds.
gi|47680918|AY586434| Influenza A Virus
(A/turkey/Italy/214845/02(H7N3)) PA gene, partial cds.
gi|47834370|AY616764| Influenza A virus (A/chicken/British
Columbia/04(H7N3)) polymerase acidic protein 2 (PA) gene, complete
cds. gi|50542647|AY646083| Influenza A virus (A/chicken/British
Columbia/GSC_human_B/04(H7N3)) polymerase acidic protein 2 (PA)
gene, complete cds. gi|50083049|AY648292| Influenza A virus
(A/GSC_chicken_B/British Columbia/04(H7N3)) polymerase acidic
protein 2 (PA) gene, complete cds. gi|50059192|AY650275| Influenza
A virus (A/GSC_chicken/British Columbia/04(H7N3)) polymerase acidic
protein 2 (PA) gene, complete cds. gi|9988639|AF268109| Influenza A
virus (A/RedKnot/Delaware/259/94(H7N7)) polymerase protein PA gene,
partial cds. gi|40353080|AJ619677| Influenza A virus
(A/chicken/Germany/R28/03(H7N7)) PA gene for polymerase complex
subunit PA, genomic RNA. gi|37813167|AY342415| Influenza A virus
(A/Netherlands/124/03(H7N7)) polymerase protein A gene, partial
cds. gi|37813169|AY342416| Influenza A virus
(A/Netherlands/126/03(H7N7)) polymerase protein A gene, partial
cds. gi|37813171|AY342417| Influenza A virus
(A/Netherlands/127/03(H7N7)) polymerase protein A gene, partial
cds. gi|37813173|AY342418| Influenza A virus
(A/Netherlands/219/03(H7N7)) polymerase protein A gene, complete
cds. gi|37813175|AY342419| Influenza A virus
(A/Netherlands/033/03(H7N7)) polymerase protein A gene, complete
cds. gi|37813177|AY342420| Influenza A virus
(A/chicken/Netherlands/1/03(H7N7)) polymerase protein A gene,
complete cds. gi|13383274|AB049157| Influenza A virus
(A/parakeet/Chiba/1/97(H9N2)) PA gene for polymerase acidic
protein, complete cds. gi|13383276|AB049158| Influenza A virus
(A/parakeet/Narita/92A/98(H9N2)) PA gene for polymerase acidic
protein, complete cds. gi|33318154|AF508662| Influenza A virus
(A/Ostrich/South Africa/9508103/95(H9N2)) segment 3 polymerase PA
(PA) gene, complete cds. gi|33318156|AF508663| Influenza A virus
(A/Chicken/Pakistan/4/99(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318158|AF508664| Influenza A virus
(A/Chicken/Pakistan/5/99(H9N2)) segment 3 polymerase PA (PA) gene,
partial cds. gi|33318160|AF508665| Influenza A virus
(A/Chicken/Germany/R45/98(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318162|AF508666| Influenza A virus
(A/Duck/Germany/113/95(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318164|AF508667| Influenza A virus
(A/Chicken/Iran/11T/99(H9N2)) segment 3 polymerase PA (PA) gene,
partial cds. gi|33318166|AF508668| Influenza A virus
(A/Chicken/Saudi Arabia/532/99(H9N2)) segment 3 polymerase PA (PA)
gene, complete cds. gi|33318168|AF508669| Influenza A virus
(A/Pheasant/Ireland/PV18/97(H9N2)) segment 3 polymerase PA (PA)
gene, complete cds. gi|33318170|AF508670| Influenza A virus
(A/Chicken/Korea/99029/99(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318172|AF508671| Influenza A virus
(A/Chicken/Beijing/8/98(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318174|AF508672| Influenza A virus
(A/Chicken/Guangdong/10/00(H9N2)) segment 3 polymerase PA (PA)
gene, complete cds. gi|33318176|AF508673| Influenza A virus
(A/Chicken/Guangdong/11/97(H9N2)) segment 3 polymerase PA (PA)
gene, complete cds. gi|33318178|AF508674| Influenza A virus
(A/Chicken/Hebei/4/98(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318180|AF508675| Influenza A virus
(A/Chicken/Heilongjiang/10/97(H9N2)) segment 3 polymerase PA (PA)
gene, complete cds. gi|33318182|AF508676| Influenza A virus
(A/Chicken/Henan/62/00(H9N2)) segment 3 polymerase PA (PA) gene,
partial cds. gi|33318184|AF508677| Influenza A virus
(A/Chicken/Ningxia/5/99(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318186|AF508678| Influenza A virus
(A/Chicken/Sichuan/5/97(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318188|AF508679| Influenza A virus
(A/Chicken/Shandong/6/96(H9N2)) segment 3 polymerase PA (PA) gene,
partial cds. gi|33318190|AF508680| Influenza A virus
(A/Chicken/Shijiazhuang/2/99(H9N2)) segment 3 polymerase PA (PA)
gene, partial cds. gi|33318192|AF508681| Influenza A virus
(A/Chicken/Shenzhen/9/97(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318194|AF508682| Influenza A virus
(A/Duck/Nanjing/1/97(H9N2)) segment 3 polymerase PA (PA) gene,
complete cds. gi|33318196|AF508683| Influenza A virus
(A/Quail/Shanghai/8/96(H9N2)) segment 3 polymerase PA (PA) gene,
partial cds. gi|31339541|AF523446| Influenza A virus
(A/Duck/Shantou/1043/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339543|AF523447| Influenza A virus
(A/Duck/Shantou/1042/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339545|AF523448| Influenza A virus
(A/Duck/Shantou/2088/01(H9N2)) polymerase (PA) gene, partial cds.
gi|31339547|AF523449| Influenza A virus
(A/Duck/Shantou/830/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339549|AF523450| Influenza A virus
(A/Duck/Shantou/1796/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339551|AF523451| Influenza A virus
(A/Duck/Shantou/2143/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339553|AF523452| Influenza A virus
(A/Duck/Shantou/2134/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339555|AF523453| Influenza A virus
(A/Duck/Shantou/2144/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339557|AF523454| Influenza A virus (A/Wild
Duck/Shantou/4808/01(H9N2)) polymerase (PA) gene, partial cds.
gi|31339559|AF523455| Influenza A virus
(A/Duck/Shantou/1881/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339561|AF523456| Influenza A virus
(A/Duck/Shantou/2102/00(H9N2)) polymerase (PA) gene, partial cds.
gi|31339563|AF523457| Influenza A virus (A/Duck/Hong
Kong/289/78(H9N2)) polymerase (PA) gene, partial cds.
gi|31339565|AF523458| Influenza A virus (A/Duck/Hong
Kong/610/79(H9N2)) polymerase (PA) gene, partial cds.
gi|31339567|AF523459| Influenza A virus (A/Duck/Hong
Kong/86/76(H9N2)) polymerase (PA) gene, partial cds.
gi|31339569|AF523460| Influenza A virus (A/Duck/Hong
Kong/366/78(H9N2)) polymerase (PA) gene, partial cds.
gi|22759040|AF536669| Influenza A virus
(A/Chicken/Beijing/1/95(H9N2)) PA gene, partial cds.
gi|22759042|AF536670| Influenza A virus
(A/Chicken/Beijing/2/97(H9N2)) PA gene, partial cds.
gi|22759044|AF536671| Influenza A virus
(A/Chicken/Beijing/3/99(H9N2)) PA gene, partial cds.
gi|22759046|AF536672| Influenza A virus
(A/Chicken/Guangdong/97(H9N2)) PA gene, partial cds.
gi|22759048|AF536673| Influenza A virus
(A/Chicken/Hebei/1/96(H9N2)) PA gene, partial cds.
gi|22759050|AF536674| Influenza A virus
(A/Chicken/Hebei/2/98(H9N2)) PA gene, partial cds.
gi|22759052|AF536675| Influenza A virus
(A/Chicken/Hebei/3/98(H9N2)) PA gene, partial cds.
gi|22759054|AF536676| Influenza A virus (A/Chicken/Henan/98(H9N2))
PA gene, partial cds. gi|22759056|AF536677| Influenza A virus
(A/Chicken/Liaoning/99(H9N2)) PA gene, partial cds.
gi|22759058|AF536678| Influenza A virus
(A/Chicken/Shandong/98(H9N2)) PA gene, partial cds.
gi|12038897|AJ291397| Influenza A virus (A/Chicken/Pakistan/2/99
(H9N2)) PA gene for polymerase PA, genomic RNA.
gi|18496121|AJ427863| Influenza A virus (A/quail/Hong Kong/FY298/00
(H9N2)) partial pa gene for PA polymerase protein, genomic RNA
gi|27465911|AY180650| Influenza A virus strain
A/Duck/Nanchang/11-392/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27465913|AY180651| Influenza A virus strain
A/Duck/Nanchang/11-290/2000
(H9N2) polymerase subunit PA (PA) gene, partial cds.
gi|27465915|AY180652| Influenza A virus strain
A/Chicken/Nanchang/1- 0016/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27465917|AY180653| Influenza A virus strain
A/Duck/Nanchang/11-197/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27465937|AY180663| Influenza A virus strain
A/Pigeon/Nanchang/2- 0461/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27465941|AY180665| Influenza A virus strain
A/Chicken/Nanchang/4- 301/2001 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27465943|AY180666| Influenza A virus strain
A/Chicken/Nanchang/4- 361/2001 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27465961|AY180675| Influenza A virus strain
A/Wild Duck/Nanchang/2- 0480/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27465983|AY180686| Influenza A virus strain
A/Duck/Nanchang/1-0070/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27465989|AY180689| Influenza A virus strain
A/Duck/Nanchang/10-389/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27465993|AY180691| Influenza A virus strain
A/Pigeon/Nanchang/11- 145/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27466001|AY180695| Influenza A virus strain
A/Quail/Nanchang/2-0460/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27466003|AY180696| Influenza A virus strain
A/Quail/Nanchang/4-040/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27466009|AY180699| Influenza A virus strain
A/Chicken/Nanchang/4- 010/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|27466015|AY180702| Influenza A virus strain
A/Duck/Nanchang/7-092/2000 (H9N2) polymerase subunit PA (PA) gene,
partial cds. gi|27466019|AY180704| Influenza A virus strain
A/Pigeon/Nanchang/7-058/2000 (H9N2) polymerase subunit PA (PA)
gene, partial cds. gi|30025973|AY253752| Influenza A virus
(A/Chicken/Shanghai/F/98(H9N2)) polymerase acidic protein (PA)
gene, complete cds. gi|49357051|AY633169| Influenza A virus
(A/mallard/Alberta/17/91(H9N2)) polymerase protein A (PA) gene,
partial cds. gi|49357079|AY633281| Influenza A virus
(A/mallard/Alberta/321/88(H9N2)) polymerase protein A (PA) gene,
partial cds. gi|49357083|AY633297| Influenza A virus
(A/mallard/Alberta/11/91(H9N2)) polymerase protein A (PA) gene,
partial cds. gi|54301528|AY664755| Influenza A virus
(A/chicken/HongKong/CSW153/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301530|AY664756| Influenza A virus
(A/chicken/HongKong/AP45/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301532|AY664757| Influenza A virus
(A/chicken/HongKong/BD90/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301534|AY664758| Influenza A virus
(A/chicken/HongKong/CSW291/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301536|AY664759| Influenza A virus
(A/chicken/HongKong/CSW304/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301538|AY664760| Influenza A virus
(A/chicken/HongKong/FY23/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301540|AY664761| Influenza A virus
(A/guineafowl/HongKong/NT101/03(H9N2)) polymerase acidic protein
(PA) gene, partial cds. gi|54301542|AY664762| Influenza A virus
(A/chicken/HongKong/NT142/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301544|AY664763| Influenza A virus
(A/chicken/HongKong/SF1/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301546|AY664764| Influenza A virus
(A/chicken/HongKong/SSP101/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301548|AY664765| Influenza A virus
(A/chicken/HongKong/TP38/03(H9N2)) polymerase acidic protein-like
(PA) gene, complete sequence. gi|54301549|AY664766| Influenza A
virus (A/chicken/HongKong/WF126/03(H9N2)) polymerase acidic protein
(PA) gene, partial cds. gi|54301551|AY664767| Influenza A virus
(A/pigeon/HongKong/WF53/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301553|AY664768| Influenza A virus
(A/pheasant/HongKong/WF54/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301555|AY664769| Influenza A virus
(A/guineafowl/HongKong/NT184/03(H9N2)) polymerase acidic protein
(PA) gene, partial cds. gi|54301557|AY664770| Influenza A virus
(A/chicken/HongKong/WF120/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301559|AY664771| Influenza A virus
(A/chicken/HongKong/NT366/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301561|AY664772| Influenza A virus
(A/chicken/HongKong/SSP418/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|54301563|AY664773| Influenza A virus
(A/chicken/HongKong/YU427/03(H9N2)) polymerase acidic protein (PA)
gene, partial cds. gi|55793682|AY800238| Influenza A virus
(A/chicken/Korea/S1/2003(H9N2)) polymerase acidic protein (PA)
gene, complete cds. gi|58429718|AY862678| Influenza A virus
(A/silky chicken/Korea/S3/03(H9N2)) PA (PA) gene, partial cds.
gi|58429720|AY862679| Influenza A virus
(A/chicken/Korea/S4/03(H9N2)) PA (PA) gene, partial cds.
gi|58429722|AY862680| Influenza A virus
(A/chicken/Korea/S5/03(H9N2)) PA (PA) gene, complete cds.
gi|58429724|AY862681| Influenza A virus
(A/chicken/Korea/S12/03(H9N2)) PA (PA) gene, partial cds.
gi|58429726|AY862682| Influenza A virus (A/duck/Korea/S13/03(H9N2))
PA (PA) gene, partial cds. gi|58429728|AY862683| Influenza A virus
(A/dove/Korea/S14/03(H9N2)) PA (PA) gene, partial cds.
gi|58429730|AY862684| Influenza A virus
(A/chicken/Korea/S15/03(H9N2)) PA (PA) gene, partial cds.
gi|58429732|AY862685| Influenza A virus
(A/chicken/Korea/S16/03(H9N2)) PA (PA) gene, partial cds.
gi|58429734|AY862686| Influenza A virus
(A/chicken/Korea/S18/03(H9N2)) PA (PA) gene, partial cds.
gi|5732356|AF156444| Influenza A virus (A/Chicken/Hong
Kong/G9/97(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732358|AF156445| Influenza A virus (A/Chicken/Hong
Kong/G23/97(H9N2)) segment 3 polymerase (PA) gene, complete cds.
gi|5732360|AF156446| Influenza A virus (A/Pigeon/Hong
Kong/Y233/97(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732362|AF156447| Influenza A virus (A/Duck/Hong
Kong/Y280/97(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732364|AF156448| Influenza A virus (A/Duck/Hong
Kong/Y439/97(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732366|AF156449| Influenza A virus (A/Quail/Hong Kong/G1/97
(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732368|AF156450| Influenza A virus (A/Chicken/Hong
Kong/739/94(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732370|AF156451| Influenza A virus (A/Quail/Hong
Kong/AF157/92(H9N2)) segment 3 polymerase (PA) gene, partial cds.
gi|5732372|AF156452| Influenza A virus
(A/Chicken/Beijing/1/94(H9N2)) segment 3 polymerase (PA) gene,
partial cds. gi|5732374|AF156453| Influenza A virus
(A/Chicken/Korea/38349- p96323/96(H9N2)) segment 3 polymerase (PA)
gene, partial cds. gi|5732376|AF156454| Influenza A virus
(A/Chicken/Korea/25232- 96006/96(H9N2)) segment 3 polymerase (PA)
gene, partial cds. gi|5732378|AF156455| Influenza A virus
(A/Shorebird/Delaware/9/96(H9N2)) segment 3 polymerase (PA) gene,
partial cds. gi|5732380|AF156456| Influenza A virus
(A/Quail/Arkansas/29209-1/93(H9N2)) segment 3 polymerase (PA) gene,
partial cds. gi|5732382|AF156457| Influenza A virus
(A/Turkey/California/189/66(H9N2)) segment 3 polymerase (PA) gene,
partial cds. gi|12060671|AF222642| Influenza A virus (A/Quail/Hong
Kong/A17/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060673|AF222643| Influenza A virus (A/Pigeon/Hong
Kong/FY6/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060675|AF222644| Influenza A virus (A/Chicken/Hong
Kong/NT16/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060677|AF222645| Influenza A virus (A/Quail/Hong
Kong/SSP10/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060679|AF222646| Influenza A virus (A/Pheasant/Hong
Kong/SSP11/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060681|AF222647| Influenza A virus (A/Chicken/Hong
Kong/FY20/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060683|AF222648| Influenza A virus (A/Chicken/Hong
Kong/KC12/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060685|AF222649| Influenza A virus (A/Quail/Hong
Kong/NT28/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060687|AF222650| Influenza A virus (A/Chicken/Hong
Kong/SF2/99(H9N2)) segment 3 PA (PA) gene, partial cds.
gi|12060689|AF222651| Influenza A virus (A/Silky Chicken/Hong
Kong/SF44/99(H9N2)) segment 3 PA (PA) gene, partial cds.
[0321]
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20070197460A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20070197460A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References