U.S. patent application number 14/929366 was filed with the patent office on 2016-05-12 for messenger una molecules and uses thereof.
The applicant listed for this patent is Arcturus Therapeutics, Inc.. Invention is credited to Padmanabh Chivukula, Joseph E. Payne, Kiyoshi Tachikawa, Luigi Warren.
Application Number | 20160130567 14/929366 |
Document ID | / |
Family ID | 55858532 |
Filed Date | 2016-05-12 |
United States Patent
Application |
20160130567 |
Kind Code |
A1 |
Chivukula; Padmanabh ; et
al. |
May 12, 2016 |
MESSENGER UNA MOLECULES AND USES THEREOF
Abstract
This invention provides a range of translatable messenger UNA
(mUNA) molecules. The mUNA molecules can be translated in vitro and
in vivo to provide an active polypeptide or protein, or to provide
an immunization agent or vaccine component. The mUNA molecules can
be used as an active agent to express an active polypeptide or
protein in cells or subjects. Among other things, the mUNA
molecules are useful in methods for treating rare diseases.
Inventors: |
Chivukula; Padmanabh; (San
Diego, CA) ; Warren; Luigi; (San Diego, CA) ;
Tachikawa; Kiyoshi; (San Diego, CA) ; Payne; Joseph
E.; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Arcturus Therapeutics, Inc. |
San Diego |
CA |
US |
|
|
Family ID: |
55858532 |
Appl. No.: |
14/929366 |
Filed: |
November 1, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62074046 |
Nov 2, 2014 |
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Current U.S.
Class: |
424/450 ;
424/208.1; 424/272.1; 424/277.1; 435/69.1; 435/69.3; 435/69.4;
514/44R; 536/23.5; 536/23.51; 536/23.7; 536/23.72 |
Current CPC
Class: |
C07K 14/4717 20130101;
A61P 1/16 20180101; C12N 2840/105 20130101; C07K 14/745 20130101;
A61P 37/04 20180101; C07K 14/505 20130101; C07K 14/575 20130101;
C12Y 304/21022 20130101; C07K 14/463 20130101; C12N 9/1018
20130101; A61K 39/00 20130101; C07K 14/8125 20130101; C12Y
201/03003 20130101; C12Y 114/16001 20130101; C12Y 204/01025
20130101; C07K 14/524 20130101; C12N 2840/60 20130101; A61K 48/00
20130101; C12N 9/1051 20130101; C12N 9/2451 20130101; C07K 14/705
20130101; C12N 9/1205 20130101; C07K 14/47 20130101; C12N 9/0071
20130101; C07K 14/805 20130101; C12N 2740/16222 20130101; C12Y
302/01033 20130101; C12N 9/88 20130101; C12N 9/644 20130101; C07K
14/445 20130101; C07K 14/4712 20130101; C12Y 207/0104 20130101;
A61P 35/00 20180101; A61P 43/00 20180101; C12Y 403/02001
20130101 |
International
Class: |
C12N 9/10 20060101
C12N009/10; C07K 14/505 20060101 C07K014/505; C07K 14/81 20060101
C07K014/81; C07K 14/705 20060101 C07K014/705; C12N 9/88 20060101
C12N009/88; C12N 9/24 20060101 C12N009/24; C07K 14/575 20060101
C07K014/575; C07K 14/47 20060101 C07K014/47; C12N 9/12 20060101
C12N009/12; C07K 14/445 20060101 C07K014/445; C07K 14/005 20060101
C07K014/005; C12N 9/64 20060101 C12N009/64; C12N 9/02 20060101
C12N009/02 |
Claims
1. A mUNA molecule, comprising one or more UNA monomers, and
comprising nucleic acid monomers, wherein the mUNA molecule is
translatable to express a polypeptide or protein.
2. The molecule of claim 1, wherein the molecule comprises from 200
to 12,000 monomers.
3. The molecule of claim 1, wherein the molecule comprises from 200
to 4,000 monomers.
4. The molecule of claim 1, wherein the molecule comprises from 1
to 8,000 UNA monomers.
5. The molecule of claim 1, wherein the molecule comprises from 1
to 100 UNA monomers.
6. The molecule of claim 1, wherein the molecule comprises from 1
to 20 UNA monomers.
7. The molecule of claim 1, wherein the molecule comprises one or
more modified nucleic acid nucleotides, or one or more
chemically-modified nucleic acid nucleotides.
8. The molecule of claim 1, wherein the molecule comprises a 5'
cap, a 5' untranslated region of monomers, a coding region of
monomers, a 3' untranslated region of monomers, and a tail region
of monomers.
9. The molecule of claim 8, wherein the molecule comprises a
translation enhancer in a 5' or 3' untranslated region.
10. The molecule of claim 1, wherein the molecule is translatable
in vivo.
11. The molecule of claim 1, wherein the molecule is translatable
in vitro.
12. The molecule of claim 1, wherein the molecule is translatable
in a mammalian cell.
13. The molecule of claim 1, wherein the molecule is translatable
in a human in vivo.
14. The molecule of claim 1, wherein a translation product of the
molecule is an active peptide or protein.
15. The molecule of claim 1, wherein a translation product of the
molecule is human EPO, human Factor IX, human alpha-1-antitrypsin,
human CFTR, human ASL, human PAH, human NIS, or human hepcidin.
16. The molecule of claim 1, wherein the molecule exhibits at least
2-fold increased translation efficiency in vivo as compared to a
native mRNA that encodes the same translation product.
17. The molecule of claim 1, wherein the molecule exhibits at least
3-fold increased translation efficiency in vivo as compared to a
native mRNA that encodes the same translation product.
18. The molecule of claim 1, wherein the molecule exhibits at least
5-fold increased translation efficiency in vivo as compared to a
native mRNA that encodes the same translation product.
19. The molecule of claim 1, wherein the molecule exhibits at least
10-fold increased translation efficiency in vivo as compared to a
native mRNA that encodes the same translation product.
20. The molecule of claim 1, wherein the molecule has a cytoplasmic
half-life in a cell at least 2-fold greater than a native mRNA of
the cell that encodes the same translation product.
21. The molecule of claim 1, wherein the molecule is a therapeutic
agent for a rare disease, a liver disease, or a cancer.
22. The molecule of claim 1, wherein the molecule is an
immunization agent or vaccine component for a rare disease, a liver
disease, or a cancer.
23. The molecule of claim 1, wherein the molecule comprises a
sequence selected from SEQ ID NOs: 1-164.
24. A composition comprising a mUNA molecule of claim 1 and a
pharmaceutically acceptable carrier.
25. A vaccine or immunization composition comprising a mUNA
molecule of claim 1.
26. The composition of claim 24, wherein the carrier is a
nanoparticle or liposome.
27. A method for ameliorating, preventing or treating a disease or
condition in a subject comprising administering to the subject a
composition of claim 24.
28. The method of claim 27, wherein the disease or condition is a
rare disease, liver disease, or cancer.
29. The method of claim 27, wherein the disease or condition is
described in Table 1.
30. A method for producing a polypeptide or protein in vivo, the
method comprising administering to a mammal a composition of claim
24.
31. The method of claim 30, wherein the polypeptide or protein is
deficient in a disease or condition described in Table 1.
32. The method of claim 30, wherein the protein is human EPO, human
Factor IX, human alpha-1-antitrypsin, human CFTR, human ASL, human
PAH, human NIS, or human hepcidin.
33. A method for producing a polypeptide or protein in vitro, the
method comprising transfecting a cell with a mUNA molecule of claim
1.
34. The method of claim 33, wherein the transfecting is done with a
transfection reagent.
35. The method of claim 33, wherein the polypeptide or protein is
deficient in a disease or condition described in Table 1.
36. The method of claim 33, wherein the protein is human EPO, human
Factor IX, human alpha-1-antitrypsin, human CFTR, human ASL, human
PAH, human NIS, or human hepcidin.
37. A method for ameliorating, preventing or treating a rare
disease or condition in a subject associated with a deficiency in a
peptide or protein, the method comprising administering to the
subject a mUNA molecule encoding the peptide or protein.
38. The method of claim 37, wherein the disease or condition
described in Table 1.
39. A method for producing a polypeptide or protein in vivo, the
method comprising administering to a mammal a mUNA molecule
encoding the polypeptide or protein.
40. The method of claim 39, wherein the polypeptide or protein is
human EPO, human Factor IX, human alpha-1-antitrypsin, human CFTR,
human ASL, human PAH, human NIS, or human hepcidin.
Description
BACKGROUND OF THE INVENTION
[0001] It has long been difficult to utilize messenger RNA
molecules in medicines. Synthetic mRNA can be designed with
inherent translational activity for making an active protein, which
could be used in various therapeutic strategies. However, the
expression of protein involves a number of steps that are localized
and/or regulated. Further, plentiful RNase enzymes can degrade
mRNA. Moreover, use of a synthetic mRNA requires clinical
formulation and delivery to cells. These steps of mRNA delivery,
partitioning and dynamics increase the need for stability and
longevity of the synthetic mRNA.
[0002] For efficient translation, natural mRNA transcripts
incorporate a 5' 7-methylguanosine cap and a 3' polyA tail. PolyA
binding proteins (PABPs) bind to the tail and cooperate with the 5'
cap via looping interactions to recruit the machinery of
translation. A 3' polyA tail of at least about 20 nucleotides is
needed to activate the mRNA for translation. Translational activity
can decrease to low levels in the absence of either the 5' cap or
the 3' polyA tail.
[0003] One drawback in using mRNA molecules in medicines is that
the lifetime of the molecule in the cytoplasm of mammalian cells is
relatively short. In general, ubiquitous mRNA degradation pathways
actively clear out transcripts from the mRNA pool. The principle
pathways for mRNA degradation involve deadenylation or trimming of
the 3' polyA tail by 3'-exoribonucleases and cleavage of the 5'-5'
triphosphate linkage that attaches the methylguanosine cap by a
decapping complex.
[0004] One way to increase mRNA longevity might be to increase
3'-nuclease resistance by incorporating nucleotide analogues or
chemical modifications in either the phosphodiester backbone or the
nucleotides, which are localized to the 3' end to be compatible
with enzymatic synthesis and efficient translation. A drawback of
this approach is that it may not be possible to selectively
incorporate such chemical modifications at 3' termini, or to retain
activity.
[0005] There is an urgent need for molecules, structures and
compositions having specific translational activity to provide
active peptides and proteins, both in vitro and in vivo. Such new
molecules having functional cytoplasmic half-life for producing
active peptides and proteins can yield new drug molecules,
therapeutic modalities, vaccines, and immunotherapies.
[0006] What is needed are translatable molecules that have
increased specific activity and/or lifetime over native mRNA, to be
used in methods and compositions for producing and delivering
active peptides and proteins in medicines.
BRIEF SUMMARY
[0007] This invention relates to the fields of molecular biology
and genetics, as well as to biopharmaceuticals and therapeutics
generated from translatable molecules. More particularly, this
invention relates to methods, structures and compositions for
molecules having translational activity for making active peptides
or proteins in vivo.
[0008] This invention provides methods and compositions for novel
molecules having translational activity, which can be used to
provide active peptides and proteins.
[0009] The molecules of this invention can have functional
cytoplasmic half-life for producing peptides and proteins. The
peptides and proteins can be active for therapeutic modalities, as
well as in vaccines and immunotherapies.
[0010] The molecules of this invention can be translatable
messenger molecules, which can have long half-life, particularly in
the cytoplasm of a cell. The longer duration of the translatable
messenger molecules of this invention can be significant for
providing a translation product that is active for ameliorating,
preventing or treating various diseases. The diseases can be
associated with undesirable modulation of protein concentration, or
undesirable activity of a protein.
[0011] This disclosure provides a range of structures for
translatable molecules that have increased specific activity and/or
lifetime over native mRNA. The translatable molecules of this
invention can be used in medicines, and for methods and
compositions for producing and delivering active peptides and
proteins.
[0012] Embodiments of this disclosure provide a wide range of
novel, translatable messenger molecules. The translatable messenger
molecules can contain monomers that are unlocked nucleomonomers
(UNA monomers). The long duration of translatable messenger UNA
molecules (mUNA molecules) of this invention can be useful for
providing an active peptide or protein translation product. The
mUNA molecules of this invention can be used in medicines for
ameliorating, preventing or treating disease.
[0013] The translatable mUNA molecules of this invention can be
used to provide peptides or proteins in vitro, ex vivo, or in
vivo.
[0014] The translatable mUNA molecules of this invention can
provide high-efficiency expression of virtually any protein.
[0015] In some embodiments, the mUNA molecules of this invention
have increased cytoplasmic half-life over a native, mature mRNA
that provides the same peptide or protein. The mUNA structures and
compositions of this invention can provide increased functional
half-life with respect to native, mature mRNAs.
[0016] In further aspects, a mUNA molecule of this invention can
provide increased activity as a drug providing a peptide or protein
product, as compared to a native, mature mRNA. In some embodiments,
a mUNA molecule can reduce the expected dose level that would be
required for efficacious therapy.
[0017] Additional embodiments of this invention can provide vaccine
compositions for immunization and immunotherapies using mUNA
molecules.
[0018] Embodiments of this invention include the following:
[0019] A mUNA molecule, containing one or more UNA monomers, and
containing nucleic acid monomers, wherein the mUNA molecule is
translatable to express a polypeptide or protein. The molecule may
have from 200 to 12,000 monomers, or from 200 to 4,000 monomers. In
some embodiments, the molecule can have from 1 to 8,000 UNA
monomers, or from 1 to 100 UNA monomers, or from 1 to 20 UNA
monomers.
[0020] A mUNA molecule can have one or more modified nucleic acid
nucleotides, and/or one or more chemically-modified nucleic acid
nucleotides.
[0021] In some embodiments, a mUNA molecule can contain a 5' cap, a
5' untranslated region of monomers, a coding region of monomers, a
3' untranslated region of monomers, and a tail region of monomers.
In certain embodiments, the molecule can contain a translation
enhancer in a 5' or 3' untranslated region.
[0022] The mUNA molecules of this invention can be translatable in
vivo, or in vitro, or in a mammalian cell, or in a human in vivo.
In some embodiments, a translation product of a mUNA molecule can
be an active peptide or protein.
[0023] In certain embodiments, a translation product of a mUNA
molecule is human EPO, human Factor IX, human alpha-1-antitrypsin,
human CFTR, human ASL, human PAH, human NIS, or human hepcidin.
[0024] In another aspect, a mUNA molecule can exhibit at least
2-fold, 3-fold, 5-fold, or 10-fold increased translation efficiency
in vivo as compared to a native mRNA that encodes the same
translation product.
[0025] In certain embodiments, a mUNA molecule can have a
cytoplasmic half-life in a cell at least 2-fold greater than a
native mRNA of the cell that encodes the same translation
product.
[0026] Embodiments of this invention further contemplate
therapeutic mUNA agents for a rare disease, a liver disease, or a
cancer. A mUNA molecule can be an immunization agent or vaccine
component for a rare disease, a liver disease, or a cancer.
[0027] This invention further provides compositions containing a
mUNA molecule and a pharmaceutically acceptable carrier, and
vaccine or immunization compositions containing a mUNA molecule.
The carrier can be a nanoparticle or liposome.
[0028] In additional embodiments, this invention provides methods
for ameliorating, preventing or treating a disease or condition in
a subject comprising administering to the subject a composition
containing a mUNA molecule. The disease or condition can be a rare
disease, liver disease, or cancer.
[0029] In certain embodiments, this invention provides methods for
producing a polypeptide or protein in vivo, by administering to a
mammal a composition containing a mUNA molecule. The polypeptide or
protein may be deficient in a disease or condition of a subject or
mammal. The protein can be human EPO, human Factor IX, human
alpha-1-antitrypsin, human CFTR, human ASL, human PAH, human NIS,
or human hepcidin.
[0030] This invention further provides methods for producing a
polypeptide or protein in vitro, by transfecting a cell with a mUNA
molecule. The polypeptide or protein can be deficient in a disease
or condition of a subject or mammal. The protein can be human EPO,
human Factor IX, human alpha-1-antitrypsin, human CFTR, human ASL,
human PAH, human NIS, or human hepcidin.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] FIG. 1 shows the results of expressing human Factor IX (F9)
in vivo using a translatable mUNA molecule of this invention, as
compared to expression of a native mRNA of Factor IX. FIG. 1 shows
that the translation efficiency of this mUNA molecule was doubled
as compared to the native mRNA of F9. The mUNA molecule of this
embodiment was translated in C57BL/c mouse to produce human F9.
[0032] FIG. 2 shows the results of expressing human Factor IX (F9)
in vitro using a translatable mUNA molecule of this invention, as
compared to expression of a native mRNA of Factor IX. FIG. 2 shows
that the translation efficiency of this mUNA molecule was increased
by 5-fold after 48 hours, as compared to the native mRNA of F9. The
mUNA molecule of this embodiment was traslated in mouse hepatocyte
cell line Hepa1-6 to produce human F9.
[0033] FIG. 3 shows the results of expressing human Erythropoietin
(EPO) in vitro using a translatable mUNA molecule of this
invention, as compared to expression of a native mRNA of human EPO.
FIG. 3 shows that the translation efficiency of this mUNA molecule
was increased nearly 3-fold after 48 hours, as compared to the
native mRNA of EPO. The mUNA molecule of this embodiment was
translated in mouse hepatocyte cell line Hepa1-6 to produce human
EPO.
[0034] FIG. 4 shows the results of expressing mouse Erythropoietin
(EPO) in vitro using several translatable mUNA molecules of this
invention, as compared to expression of a native mRNA of mouse EPO.
FIG. 4 shows that the translation efficiencies of the mUNA
molecules (#2, 3, 4, 5, 6, 7, 8, 9, 10 and 11) were increased by up
to 10-fold after 72 hours, as compared to the native mRNA of EPO.
The mUNA molecules of this embodiment were translated in mouse
hepatocyte cell line Hepa1-6 to produce mouse EPO.
[0035] FIG. 5 shows the results of expressing human
alpha-1-antitrypsin in vivo using a translatable mUNA molecule of
this invention, as compared to expression of a native mRNA of human
alpha-1-antitrypsin. FIG. 5 shows that the translation efficiency
of this mUNA molecule at 72 hrs was increased more than 3-fold as
compared to the native mRNA of human alpha-1-antitrypsin. The mUNA
molecule of this embodiment was translated in C57BL/c mouse to
produce human alpha-1-antitrypsin.
[0036] FIG. 6 shows the results of expressing human erythropoietin
(EPO) in vivo using a translatable mUNA molecule of this invention,
as compared to expression of a native mRNA of human EPO. FIG. 6
shows that the translation efficiency of this mUNA molecule at 72
hrs was increased more than 10-fold as compared to the native mRNA
of human EPO. The mUNA molecule of this embodiment was translated
in C57BL/c mouse to produce human EPO.
[0037] FIG. 7 shows the primary structure of a functional mRNA
transcript in the cytoplasm. The mRNA includes a 5' methylguanosine
cap, a protein coding sequence flanked by untranslated regions
(UTRs), and a polyadenosine (polyA) tail bound by polyA binding
proteins (PABPs).
[0038] FIG. 8 shows the 5' cap and PABPs cooperatively interacting
with proteins involved in translation to facilitate the recruitment
and recycling of ribosome complexes.
[0039] FIG. 9 shows the splint-mediated ligation scheme, in which
an acceptor RNA with a 30-monomer stub polyA tail (A(30)) was
ligated to a 30-monomer donor oligomer A(30). The splint-mediated
ligation used a DNA oligomer splint which was complementary to the
3' UTR sequence upstream of the stub polyA tail, and included a
60-monomer oligo(dT) 5' heel (T(60)) to splint the ligation. The
anchoring region of the splint was complementary to the UTR
sequence to ensure that a 5' dT.sub.30 overhang was presented upon
hybridization to the acceptor. This brings the donor oligomer into
juxtaposition with the 3' terminus of the stub tail, dramatically
improving the kinetics of ligation.
[0040] FIG. 10 shows experimental results of splint-mediated
ligation of a donor oligomer to an acceptor. FIG. 10 shows the
results of ligation using 2 ug of a 120-monomer acceptor with an
A.sub.30 stub tail that was ligated to a 5'-phosphorylated A.sub.30
RNA donor oligomer using T4 RNA Ligase 2. The reaction was
incubated overnight at 37.degree. C. The ligation and a mock
reaction done without enzyme were purified, treated with DNAse I
for 1 hour to degrade and detach the splint oligomers, and
re-purified in a volume of 30 uL. The ligation efficiency was
nearly 100%. The absence of a size shift in the mock-reaction prep
shows that the acceptor and donor were truly ligated and not simply
held together by undigested splint oligomers.
[0041] FIG. 11 shows the results of splint-mediated ligation using
an acceptor RNA with a 30-monomer stub polyA tail (A(30)). The
ligation reactions were performed with three different donor
oligomer species: A(30), A(60), and A(120). Based on the gel
shifts, the ligations have attained nearly 100% efficiency.
[0042] FIG. 12 shows the results of one-hour splint-mediated
ligations that were performed on nGFP-A.sub.30 transcripts. The
resulting ligation products were compared to untreated transcripts
and native nGFP-A.sub.60 IVT products. The native nGFP-A.sub.60 and
the ligated products were up-shifted on the gel relative to the
untreated nGFP-A.sub.30 transcripts and mock-ligated material,
showing that the ligation yield was nearly 100%.
[0043] FIG. 13 shows increased lifetime and translational activity
for an nGFP-A.sub.60 ligation product. In FIG. 13, nuclearized
transcripts were transfected into fibroblasts, and a comparison of
fluoresence signals was made for nGFP-A.sub.30, mock-ligated
nGFP-A.sub.30, and an nGFP-A.sub.60 ligation product (FIG. 13, left
to right). The significantly higher fluorescence signal observed
for the nGFP-A.sub.60 ligation product shows that it has markedly
increased translational activity.
[0044] FIG. 14 shows the results of a ligation performed with a
100-monomer acceptor RNA that was treated for 3 hours at room
temperature with T4 RNA Ligase 2 (truncated KQ mutant) using a 10
uM concentration of a polyA tail 30-monomer donor oligomer. 15% PEG
8000 was included in the reaction as a volume excluder to promote
efficient ligation. The ligation reaction showed that a high
molecular weight product was formed, having a size in between the
100-monomer acceptor RNA and a 180-monomer RNA transcript included
as a size standard. These results show that the ligation reaction
produced a predominant product having high molecular weight with
nearly 100% ligation of the donor to the acceptor. Additional
experiments with concentrations of the polyA tail at 10 uM, 20 uM,
and 40 uM showed that from about 50% to about 100% of the acceptor
RNA was ligated.
DETAILED DESCRIPTION OF THE INVENTION
[0045] This invention provides a range of novel agents and
compositions to be used for therapeutic applications. The molecules
and compositions of this invention can be used for ameliorating,
preventing or treating various diseases associated with genomic
functionalities.
[0046] The molecules of this invention can be translatable
messenger UNA molecules, which can have long half-life,
particularly in the cytoplasm. The long duration mUNA molecules
(mUNA molecules) can be used for ameliorating, preventing or
treating various diseases associated with undesirable modulation of
protein concentration, or activity of a protein.
[0047] The properties of the mUNA compounds of this invention arise
according to their molecular structure, and the structure of the
molecule in its entirety, as a whole, can provide significant
benefits based on those properties. Embodiments of this invention
can provide mUNA molecules having one or more properties that
advantageously provide enhanced effectiveness in regulating protein
expression or concentration, or modulating protein activity. The
molecules and compositions of this invention can provide
formulations for therapeutic agents for various diseases and
conditions, which can provide clinical agents.
[0048] This invention provides a range of mUNA molecules that are
surprisingly translatable to provide active peptide or protein, in
vitro and in vivo.
[0049] The mUNA structures and compositions can have increased
translational activity and cytoplasmic half-life. In these
embodiments, the mUNA structures and compositions can provide
increased functional half-life in the cytoplasm of mammalian cells
over native mRNA molecules. The inventive mUNA molecules can have
increased half-life of activity with respect to a corresponding
native mRNA.
[0050] A wide range of novel mUNA molecules are provided herein,
each of which can incorporate specialized linker groups. The linker
groups can be attached in a chain in the mUNA molecule. Each linker
group can also be attached to a nucleobase.
[0051] In some aspects, a linker group can be a monomer. Monomers
can be attached to form a chain molecule. In a chain molecule of
this invention, a linker group monomer can be attached at any point
in the chain.
[0052] In certain aspects, linker group monomers can be attached in
a chain molecule of this invention so that the linker group
monomers reside near the ends of the chain, or at any position in
the chain.
[0053] As used herein, a chain molecule can also be referred to as
an oligomer.
[0054] In further aspects, the linker groups of a chain molecule
can each be attached to a nucleobase. The presence of nucleobases
in the chain molecule can provide a sequence of nucleobases in the
chain molecule.
[0055] In certain embodiments, this invention provides oligomer
mUNA molecules having chain structures that incorporate novel
combinations of the linker group monomers, along with certain
natural nucleotides, or non-natural nucleotides, or modified
nucleotides, or chemically-modified nucleotides.
[0056] The oligomer mUNA molecules of this invention can display a
sequence of nucleobases, and can be designed to express a
polypeptide or protein, in vitro, ex vivo, or in vivo. The
expressed polypeptide or protein can have activity in various
forms, including activity corresponding to protein expressed from
natural mRNA, or activity corresponding to a negative or dominant
negative protein.
[0057] In some aspects, this invention can provide active mUNA
oligomer molecules having a base sequence that corresponds to at
least a fragment of a native nucleic acid molecule of a cell.
[0058] In some embodiments, the cell can be a eukaryotic cell, a
mammalian cell, or a human cell.
[0059] This invention provides structures, methods and compositions
for oligomeric mUNA agents that incorporate the linker group
monomers. The oligomeric molecules of this invention can be used as
active agents in formulations for therapeutics.
[0060] This invention provides a range of mUNA molecules that are
useful for providing therapeutic effects because of their longevity
of activity in providing an expressed peptide or protein.
[0061] In certain embodiments, an active mUNA molecule can be
structured as an oligomer composed of monomers. The oligomeric
structures of this invention may contain one or more linker group
monomers, along with certain nucleotides.
[0062] An expressed peptide or protein can be modified or mutated
as compared to a native variant, or can be a homolog or ortholog
for enhanced expression in a eukaryotic cell. An active mUNA
molecule can be human codon optimized. Methodologies for optimizing
codons are known in the art.
[0063] In certain embodiments, a mUNA molecule may contain a
sequence of nucleobases, and can be designed to express a peptide
or protein of any isoform, in part by having sufficient homology
with a native polynucleotide sequence.
[0064] In some embodiments, a mUNA molecule can be from about 200
to about 12,000 monomers in length, or more. In certain
embodiments, a mUNA molecule can be from 200 to 12,000 monomers in
length, or 200 to 10,000 monomers, or 200 to 8,000 monomers, or 200
to 6000 monomers, or 200 to 5000 monomers, or 200 to 4000 monomers,
or 200 to 3600 monomers, or 200 to 3200 monomers, or 200 to 3000
monomers, or 200 to 2800 monomers, or 200 to 2600 monomers, or 200
to 2400 monomers, or 200 to 2200 monomers, or 600 to 3200 monomers,
or 600 to 3000 monomers, or 600 to 2600 monomers.
[0065] In some embodiments, a mUNA molecule can contain from 1 to
about 8,000 UNA monomers. In certain embodiments, a mUNA molecule
can contain from 1 to 8,000 UNA monomers, or 1 to 6,000 UNA
monomers, or 1 to 4,000 UNA monomers, or 1 to 3,000 UNA monomers,
or 1 to 2,000 UNA monomers, or 1 to 1,000 UNA monomers, or 1 to 500
UNA monomers, or 1 to 300 UNA monomers, or 1 to 200 UNA monomers,
or 1 to 100 UNA monomers, or 1 to 50 UNA monomers, or 1 to 40 UNA
monomers, or 1 to 30 UNA monomers, or 1 to 20 UNA monomers, or 1 to
10 UNA monomers, or 1 to 6 UNA monomers.
[0066] In some embodiments, a mUNA molecule can be from about 200
to about 12,000 bases in length, or more. In certain embodiments, a
mUNA molecule can be from 200 to 12,000 bases in length, or 200 to
10,000 bases, or 200 to 8,000 bases, or 200 to 6000 bases, or 200
to 5000 bases, or 200 to 4000 bases, or 200 to 3600 bases, or 200
to 3200 bases, or 200 to 3000 bases, or 200 to 2800 bases, or 200
to 2600 bases, or 200 to 2400 bases, or 200 to 2200 bases, or 600
to 3200 bases, or 600 to 3000 bases, or 600 to 2600 bases.
[0067] A mUNA molecule of this invention may comprise a 5' cap, a
5' untranslated region of monomers, a coding region of monomers, a
3' untranslated region of monomers, and a tail region of monomers.
Any of these regions of monomers may comprise one or more UNA
monomers.
[0068] A mUNA molecule of this invention may comprise a 5'
untranslated region of monomers containing one or more UNA
monomers.
[0069] A mUNA molecule of this invention may comprise a coding
region of monomers containing one or more UNA monomers.
[0070] A mUNA molecule of this invention may comprise a 3'
untranslated region of monomers containing one or more UNA
monomers.
[0071] A mUNA molecule of this invention may comprise a tail region
of monomers containing one or more UNA monomers.
[0072] A mUNA molecule of this invention may comprise a 5' cap
containing one or more UNA monomers.
[0073] A mUNA molecule of this invention can be translatable, and
may comprise regions of sequences or structures that are operable
for translation in a cell, or which have the functionality of
regions of an mRNA including, for example, a 5' cap, a 5'
untranslated region, a coding region, a 3' untranslated region, and
a polyA tail.
[0074] This invention further contemplates methods for delivering
one or more vectors, or one or more mUNA molecules to a cell.
[0075] In some embodiments, one or more mUNA molecules can be
delivered to a cell, in vitro, ex vivo, or in vivo. Viral and
non-viral transfer methods as are known in the art can be used to
introduce mUNA molecules in mammalian cells. mUNA molecules can be
delivered with a pharmaceutically acceptable vehicle, or for
example, encapsulated in a liposome.
[0076] A peptide or protein expressed by a mUNA molecule can be any
peptide or protein, endogenous or exogenous in nature with respect
to a eukaryotic cell, and may be a synthetic or non-natural peptide
or protein with activity or effect in the cell.
[0077] In some embodiments, mUNA structures and compositions of
this invention can reduce the number and frequency of transfections
required for cell-fate manipulation in culture as compared to
utilizing native compositions.
[0078] In additional aspects, this invention provides increased
activity for mUNA-based drugs as compared to utilizing native
compositions, and can reduce the dose levels required for
efficacious therapy.
[0079] In further aspects, this invention provides increased
activity for mUNA-based molecules, as compared to utilizing a
native mRNA as active agent.
[0080] In some aspects, this invention can provide mUNA molecules
that may reduce the cellular innate immune response, as compared to
that induced by a natural nucleic acid, peptide or protein.
[0081] In further aspects, embodiments of this invention can
provide increased efficacy for single-dose therapeutic modalities,
including mUNA immunization and immunotherapies.
[0082] This invention can provide synthetic mUNA molecules that are
refractory to deadenylation as compared to native molecules.
[0083] In certain embodiments, this invention can provide synthetic
mUNA molecules with increased specific activity and longer
functional half-life as compared to native molecules. The synthetic
mUNA molecules of this invention can provide increased levels of
ectopic protein expression. When using a mUNA molecule as a vector,
cellular-delivery can be at increased levels, and cytotoxic innate
immune responses can be restrained so that higher levels of ectopic
protein expression can be achieved. The mUNA molecules of this
invention can have increased specific activity and longer
functional half-life than mRNAs.
[0084] In certain aspects, a mUNA molecule may have a number of
mutations from a native mRNA, or from a disease associated
mRNA.
[0085] In further embodiments, this invention can provide mUNA
molecules having cleavable delivery and targeting moieties attached
at the 3' end.
[0086] In general, the specific activity for a synthetic
translatable molecule delivered by transfection can be viewed as
the number of molecules of protein expressed per delivered
transcript per unit time.
[0087] As used herein, translation efficiency refers to a measure
of the production of a protein or polypeptide by translation of a
messenger molecule in vitro or in vivo.
[0088] This invention provides a range of mUNA molecules, which can
contain one or more UNA monomers, and a number of nucleic acid
monomers, wherein the mUNA molecule can be translated to express a
polypeptide or protein.
[0089] In some embodiments, this invention includes a range of mUNA
molecules, which contain one or more UNA monomers in one or more
untranslated regions, and a number of nucleic acid monomers,
wherein the mUNA molecule can be translated to express a
polypeptide or protein.
[0090] In some embodiments, this invention includes a range of mUNA
molecules, which contain one or more UNA monomers in a tail region
or monomers, and a number of nucleic acid monomers, wherein the
mUNA molecule can be translated to express a polypeptide or
protein.
[0091] In some embodiments, a mUNA molecule can contain a modified
5' cap.
[0092] In some embodiments, a mUNA molecule can contain one ore
more UNA monomers in a 5' cap.
[0093] In further embodiments, a mUNA molecule can contain a
translation enhancing 5' untranslated region of monomers.
[0094] In further embodiments, a mUNA molecule can contain one or
more UNA monomers in a 5' untranslated region.
[0095] In additional embodiments, a mUNA molecule can contain a
translation enhancing 3' untranslated region of monomers.
[0096] In additional embodiments, a mUNA molecule can contain one
or more UNA monomers in a 3' untranslated region of monomers.
[0097] In additional embodiments, a mUNA molecule can contain one
or more UNA monomers in a tail region of monomers.
[0098] In additional embodiments, a mUNA molecule can contain one
or more UNA monomers in a polyA tail.
[0099] In another aspect, a mUNA molecule can exhibit at least
2-fold, 3-fold, 5-fold, or 10-fold increased translation efficiency
in vivo as compared to a native mRNA that encodes the same
translation product.
[0100] In another aspect, a mUNA molecule can produce at least
2-fold, 3-fold, 5-fold, or 10-fold increased polypeptide or protein
in vivo as compared to a native mRNA that encodes the same
polypeptide or protein.
[0101] In additional embodiments, this invention provides methods
for treating a rare disease or condition in a subject by
administering to the subject a composition containing a mUNA
molecule.
[0102] In additional embodiments, this invention provides methods
for treating a liver disease or condition in a subject by
administering to the subject a composition containing a mUNA
molecule.
[0103] Modalities for Peptides and Proteins
[0104] A mUNA molecule of this invention may be used for
ameliorating, preventing or treating a disease through enzyme
modulation or replacement. In these embodiments, a mUNA molecule of
this invention can be administered to regulate, modulate, increase,
or decrease the concentration or effectiveness of a natural enzyme
in a subject.
[0105] In some aspects, the enzyme can be an unmodified, natural
enzyme for which the patient has an abnormal quantity.
[0106] In some embodiments, a mUNA molecule can be delivered to
cells or subjects, and translated to supply increased levels of the
natural enzyme.
[0107] A mUNA molecule of this invention may be used for
ameliorating, preventing or treating a disease through modulation
or introduction of a peptide or protein. In these embodiments, a
mUNA molecule of this invention can be administered to regulate,
modulate, increase, or decrease the concentration or effectiveness
of a peptide or protein in a subject, where the peptide or protein
is non-natural or mutated, as compared to a native peptide or
protein.
[0108] In some aspects, the peptide or protein can be a modified,
non-natural, exogenous, or synthetic peptide or protein, which has
a pharmacological effect in a subject.
[0109] In some embodiments, a mUNA molecule can be delivered to
cells or subjects, and translated to supply a concentration of the
peptide or protein.
[0110] Examples of diseases for enzyme modulation include lysosomal
diseases, for example, Gaucher disease, Fabry disease,
Mucopolysaccharidoses (MPS) and related diseases including MPS I,
MPS II (Hunter syndrome), and MPS VI, as well as Glycogen storage
disease type II.
[0111] Examples of diseases for enzyme modulation include
hematologic diseases, for example, sickle-cell disease,
thalassemia, methemoglobinemia, anemia due to deficiency of
hemoglobin or B.sub.12 intrinsic factor, spherocytosis,
glucose-6-phosphate dehydrogenase deficiency, and pyruvate kinase
deficiency.
[0112] Examples of diseases for enzyme modulation include
hemophilia, Von Willebrand disease, Protein S deficiency,
age-related macular degeneration, trinucleotide repeat disorders,
muscular dystrophy, insertion mutation diseases, DNA
repair-deficiency disorders, and deletion mutation diseases.
[0113] Rare Diseases
[0114] Examples of diseases and/or conditions for which the mUNA
molecules of this invention can be translatable to provide an
active agent include those in Table 1.
TABLE-US-00001 TABLE 1 Rare diseases RARE DISEASE DEFICIENCY
Aminoacylase 1 deficiency Aminoacylase 1 Apo A-I deficiency Apo A-I
Carbamoyl phosphate synthetase 1 Carbamoyl phosphate synthetase 1
deficiency Ornithine transcarbamylase Ornithine transcarbamylase
deficiency Plasminogen activator inhibitor Plasminogen activator
inhibitor type 1 type 1 deficiency Flaujeac factor deficiency
Flaujeac factor (High-molecular-weight kininogen)
High-molecular-weight kininogen High-molecular-weight kininogen
(Flaujeac factor) deficiency congenital PEPCK 1 deficiency PEPCK 1
Pyruvate kinase deficiency liver Pyruvate kinase liver type type
Alpha 1-antitrypsin deficiency Alpha 1-antitrypsin Anti-plasmin
deficiency congenital Anti-plasmin Apolipoprotein C 2I deficiency
Apolipoprotein C 2I Butyrylcholinesterase deficiency
Butyrylcholinesterase Complement component 2 Complement component 2
deficiency Complement component 8 Complement component 8 type 2
deficiency type 2 Congenital antithrombin Antithrombin deficiency
type 1 Congenital antithrombin Antithrombin, type 2 deficiency type
2 Congenital antithrombin Antithrombin, type 3 deficiency type 3
Cortisone reductase deficiency 1 Cortisone reductase Factor VII
deficiency Factor VII Factor X deficiency Factor X Factor XI
deficiency Factor XI Factor XII deficiency Factor XII Factor XIII
deficiency Factor XIII Fibrinogen deficiency congenital Fibrinogen
Fructose-1 6-bisphosphatase Fructose-1 6-bisphosphatase deficiency
Gamma aminobutyric acid Gamma aminobutyric acid transaminase
transaminase deficiency Gamma-cystathionase deficiency
Gamma-cystathionase Glut2 deficiency Glut2 GTP cyclohydrolase I
deficiency GTP cyclohydrolase I Isolated growth hormone Isolated
growth hormone type 1B deficiency type 1B Molybdenum cofactor
deficiency Molybdenum cofactor Prekallikrein deficiency congenital
Prekallikrein Proconvertin deficiency congenital Proconvertin
Protein S deficiency Protein S Pseudocholinesterase deficiency
Pseudocholinesterase Stuart factor deficiency congenital Stuart
factor Tetrahydrobiopterin deficiency Tetrahydrobiopterin Type 1
plasminogen deficiency Plasminogen Urocanase deficiency Urocanase
Chondrodysplasia punctata with Chondrodysplasia punctata with
steroid sulfatase/X- steroid sulfatase deficiency linked
chondrodysplasia punctata 1 Homocystinuria due to CBS CBS
deficiency Guanidinoacetate Guanidinoacetate methyltransferase
methyltransferase deficiency Pulmonary surfactant protein B
Pulmonary surfactant protein B deficiency Aminoacylase 1 deficiency
Aminoacylase 1 Acid Sphingomyelinase Enzyme found in lysosomes,
responsible for conversion of Deficiency lipid sphingomyelin into
lipid ceramide Adenylosuccinate Lyase Neurological disorder, brain
dysfunction (encephalopathy) Deficiency and to delayed development
of mental and movement abilities, autistic behaviors and seizures
Aggressive Angiomyxoma Myxoid tumor involving the blood vessels,
may be a non- metastasizing benign tumor Albrights Hereditary
Inherited in an autosomal dominant pattern, lack of Osteodystrophy
responsiveness to parathyroid hormone, low serum calcium, high
serum phosphate Carney Stratakis Syndrome Very rare syndrome
characterized by gastrointestinal stromal tumors and
paragangliomas. Carney Triad Syndrome Characterized by the
coexistence of 3 types of neoplasms, mainly in young women,
including gastric gastrointestinal stromal tumor, pulmonary
chondroma, and extra-adrenal paraganglioma CDKL5 Mutation Results
in severe neurodevelopmental impairment and early onset, difficult
to control seizures CLOVES Syndrome Complex vascular anomalies:
Congenital, Lipomatous Overgrowth, Vascular malformations,
Epidermal nevi and Scoliosis/Skeletal/Spinal anomalies Cockayne
Syndrome Characterized by short stature and an appearance of
premature aging, failure to gain weight, abnormally small head
size, and impaired development of the nervous system Congenital
Disorder of Rare inborn errors of metabolism involving deficient or
Glycosylation type 1R defective glycosylation Cowden Syndrome
Characterized by multiple noncancerous, tumor-like growths called
hamartomas and an increased risk of developing certain cancers DEND
Syndrome Generally severe form of neonatal diabetes mellitus
characterized by a triad of developmental delay, epilepsy, and
neonatal diabetes Dercum's Disease Characterized by multiple, and
painful lipomas. These lipomas mainly occur on the trunk, the upper
arms and upper legs Febrile Infection-Related Epilepsy
Explosive-onset, potentially fatal acute epileptic Syndrome
encephalopathy, develops in previously healthy children and
adolescents following the onset of a non-specific febrile illness
Fibular Aplasia Tibial Campomelia Unknown genetic basis and
inheritance with variable Oligosyndactyly Syndrome expressivity and
penetrance Food Protein-Induced Enterocolitis A non-IgE mediated
immune reaction in the gastrointestinal Syndrome system to one or
more specific foods, commonly characterized by profuse vomiting and
diarrhea Foreign Body Giant Cell Reactive Collection of fused
macrophages which are generated in Tissue Disease response to the
presence of a large foreign body; particularly evident with
implants that cause the body chronic inflammation and foreign body
response Galloway-Mowat Physical features may include an unusually
small head and additional abnormalities of the head and facial
area; damage to clusters of capillaries in the kidneys resulting in
abnormal kidney function; and, in many cases, protrusion of part of
the stomach through an abnormal opening in the diaphragm Gitelman
syndrome Autosomal recessive kidney disorder characterized by
hypokalemic metabolic alkalosis with hypocalciuria, and
hypomagnesemia. Glycerol Kinase Deficiency X-linked recessive
enzyme defect that is heterozygous in nature, responsible gene in a
region containing genes in which deletions can cause DMD and
adrenal hypoplasia congenita Glycogen Storage Disease type 9 Caused
by the inability to break down glycogen. The different forms of the
condition can affect glycogen breakdown in liver cells, muscle
cells or both gm1 gangliosidosis Autosomal recessive lysosomal
storage disease characterized by accumulation of ganglioside
substrates in lysosomes Hereditary spherocytosis Affects red blood
cells, shortage of red blood cells, yellowing of the eyes and skin,
and an enlarged spleen Hidradenitis Suppurativa Stage III Disorder
of the terminal follicular epithelium in the apocrine gland-bearing
skin, frequently causing keloids, contractures, and immobility.
Stage III is defined as multiple lesions, with more extensive sinus
tracts and scarring Horizonatal Gaze Palsy with Disorder that
affects vision and also causes an abnormal Progressive Scoliosis
curvature of the spine IMAGe syndrome The combination of
intrauterine growth restriction, metaphyseal dysplasia, adrenal
hypoplasia congenita, and genital anomalies (only about 20 cases
reported in the medical literature) Isodicentric 15 Chromosome
abnormality in which a child is born with extra genetic material
from chromosome 15 isolated hemihyperplasia One side of the body
grows more than other, causing asymmetry Juvenile Xanthogranuloma
Usually benign and self-limiting. It occurs most often in the skin
of the head, neck, and trunk but can also occur in the arms, legs,
feet, and buttocks Kasabach-Merritt Syndrome A vascular tumor leads
to decreased platelet counts and sometimes other bleeding problems
Kniest Dysplasia Disorder of bone growth characterized by short
stature (dwarfism) with other skeletal abnormalities and problems
with vision and hearing Koolen de-Vries Syndrome Disorder
characterized by developmental delay and mild to moderate
intellectual disability. They usually have weak muscle tone in
childhood. About half have recurrent seizures Lennox-Gastaut
syndrome Type of epilepsy with multiple different types of
seizures, particularly tonic (stiffening) and atonic (drop)
seizures. Intellectual development is usually, but not always,
impaired Lymphangiomatosis Congenital and can affect any of the
body's systems except the central nervous system (including the
brain) Lymphangiomiomytosis Can occur either sporadically or in
association with the tuberous sclerosis complex (TSC) and is often
considered a forme fruste of TSC MASA Syndrome X-linked recessive
neurological disorder Mast Cell Activation disorder Condition with
signs and symptoms involving the skin, gastrointestinal,
cardiovascular, respiratory, and neurologic systems Mecp2
Duplication Syndrome Genetic neurodevelopmental disorder
characterized by low muscle tone, potentially severe intellectual
disability, developmental delays, recurrent respiratory infections,
speech abnormalities, seizures, and progressive spasticity Mucha
Habermann Skin disorder Neonatal Hemochromatosis Severe liver
disease of fetal or perinatal onset, associated with deposition of
stainable iron in extrahepatic sites, disordered iron handling due
to injury to the perinatal liver, as a form of fulminant hepatic
failure N-glycanase deficiency The encoded enzyme may play a role
in the proteasome- mediated degradation of misfolded glycoproteins
Opsoclonus Myoclonus Syndrome Neurological disorder of unknown
causes which appears to be the result of an autoimmune process
involving the nervous system Persistent genital arousal disorder
Results in a spontaneous, persistent, and uncontrollable genital
arousal, with or without orgasm or genital engorgement, unrelated
to any feelings of sexual desire Pompe Disease Inherited disorder
caused by the buildup of glycogen in the body's cells. The
accumulation of glycogen in certain organs and tissues, especially
muscles, impairs their ability to function normally Progressive
Familial Intrahepatic Disorder that causes progressive liver
disease, which Cholestasis typically leads to liver failure. In
people with PFIC, liver cells are less able to secrete a digestive
fluid called bile. The buildup of bile in liver cells causes liver
disease in affected individuals Pseudohypoparathyroidism type 1a
Characterized by renal resistance to parathyroid hormone, resulting
in hypocalcemia, hyperphosphatemia, and elevated PTH; resistance to
other hormones including thydroid stimulating hormone,
gonadotropins and growth- hormone-releasing hormone PTEN Hamartoma
Tumor The gene was identified as a tumor suppressor that is
Syndrome mutated in a large number of cancers at high frequency
Schnitzler syndrome Characterised by chronic hives and periodic
fever, bone pain and joint pain (sometimes with joint
inflammation), weight loss, malaise, fatigue, swollen lymph glands
and enlarged spleen and liver Scleroderma Chronic hardening and
tightening of the skin and connective tissues
Semi Lobar Holoprosencephany Holoprosencephany: birth defect of the
brain, which often can also affect facial features, including
closely spaced eyes, small head size, and sometimes clefts of the
lip and roof of the mouth. Semilobar holoprosencephaly is a subtype
of holoprosencephaly characterised by an incomplete forebrain
division Sjogren's Syndrome Immune system disorder characterized by
dry eyes and dry mouth Specific Antibody Deficiency Immune Disease
SYNGAP 1 A ras GTPase-activating protein that is critical for the
development of cognition and proper synapse function Trigeminal
Trophic Syndrome This is the wing of tissue at the end of the nose
above the nostril. Trigeminal trophic syndrome is due to damage to
the trigeminal nerve Undiffentiated Connective Tissue Systemic
autoimmune disease Disease X-linked hypophosphatemia X-linked
dominant form of rickets (or osteomalacia) that differs from most
cases of rickets in that ingestion of vitamin D is relatively
ineffective. It can cause bone deformity including short stature
and genu varum
[0115] Modalities for Immune Modulation
[0116] The mUNA molecules of this invention can be translatable to
provide an active protein. In certain embodiments, a translatable
mUNA molecule can provide an active mRNA immunization agent, or an
mRNA vaccine component.
[0117] A mUNA vaccine of this disclosure can advantageously provide
a safe and efficacious genetic vaccine by inducing an immune
response having both cellular and humoral components. In general,
protein can be expressed using a mUNA vaccine of this
invention.
[0118] In some embodiments, a mUNA vaccine can advantageously
provide protein synthesis in the cytoplasm. In certain embodiments,
a mUNA vaccine of this invention can provide internalization,
release and transport of an exogenous mRNA in the cytoplasm.
[0119] In certain aspects, a mUNA vaccine of this invention can
encode for a protein antigen that can be translated by host
cells.
[0120] In further aspects, some mUNA vaccines of this disclosure
can encode for tumor antigens, viral antigens, or allergens.
[0121] Modalities for administering a mUNA vaccine of this
invention can include intravenous, intranodal, intradermal,
subcutaneous and intrasplenic.
[0122] Embodiments of this invention further provide mUNA vaccines
having increased half-life of translation, which can be used to
reduce the necessary dose and exposure to antigen, and reduce the
risk of inducing tolerance.
[0123] A mUNA vaccine of this invention can provide an
immunological effect without the risk of integration of a component
into the genome, and may reduce the risk of mutagenesis as compared
to other genetic vaccines.
[0124] Additional embodiments of this disclosure include mUNA
molecules having translational activity, where the translational
activity can be described by a cytoplasmic half-life in a mammalian
cell. The half-life can be determined by the time required for 50%
of the mUNA molecule to be degraded in the cell.
[0125] A translatable mUNA molecule of this invention can be a
precursor of an active molecule, which can be used in the treatment
of a condition or disease in a subject.
[0126] In some embodiments, a translatable mUNA molecule of this
invention can be a pharmacologically active molecule having
increased half-life in the cytoplasm of mammalian cells.
[0127] Examples of mUNA molecules of this invention include a mUNA
molecule that provides an mRNA encoding HIV-1 gag antigen, a mUNA
molecule that provides an mRNA encoding antigens overexpressed in
lung cancers, a mUNA molecule that provides an mRNA encoding
malarial P. falciparum reticulocyte-binding protein homologue 5
(PfRHS), and a mUNA molecule that provides an mRNA encoding
malarial Plasmodium falciparum PfSEA-1, a 244 KD malaria antigen
expressed in schizont-infected RBCs.
[0128] UNA Monomers and Oligomers
[0129] In some embodiments, linker group monomers can be unlocked
nucleomonomers (UNA monomers), which are small organic molecules
based on a propane-1,2,3-tri-yl-trisoxy structure as shown
below:
##STR00001##
where R.sup.1 and R.sup.2 are H, and R.sup.1 and R.sup.2 can be
phosphodiester linkages, Base can be a nucleobase, and R.sup.3 is a
functional group described below.
[0130] In another view, the UNA monomer main atoms can be drawn in
IUPAC notation as follows:
##STR00002##
where the direction of progress of the oligomer chain is from the
1-end to the 3-end of the propane residue.
[0131] Examples of a nucleobase include uracil, thymine, cytosine,
5-methylcytosine, adenine, guanine, inosine, and natural and
non-natural nucleobase analogues.
[0132] Examples of a nucleobase include pseudouracil,
1-methylpseudouracil, and 5-methoxyuracil.
[0133] In general, a UNA monomer, which is not a nucleotide, can be
an internal linker monomer in an oligomer. An internal UNA monomer
in an oligomer is flanked by other monomers on both sides.
[0134] A UNA monomer can participate in base pairing when the
oligomer forms a complex or duplex, for example, and there are
other monomers with nucleobases in the complex or duplex.
[0135] Examples of UNA monomer as internal monomers flanked at both
the propane-1-yl position and the propane-3-yl position, where
R.sup.3 is --OH, are shown below.
##STR00003##
[0136] A UNA monomer can be a terminal monomer of an oligomer,
where the UNA monomer is attached to only one monomer at either the
propane-1-yl position or the propane-3-yl position. Because the UNA
monomers are flexible organic structures, unlike nucleotides, the
terminal UNA monomer can be a flexible terminator for the
oligomer.
[0137] Examples of a UNA monomer as a terminal monomer attached at
the propane-3-yl position are shown below.
##STR00004##
[0138] Because a UNA monomer can be a flexible molecule, a UNA
monomer as a terminal monomer can assume widely differing
conformations. An example of an energy minimized UNA monomer
conformation as a terminal monomer attached at the propane-3-yl
position is shown below.
##STR00005##
[0139] UNA-A terminal forms: the dashed bond shows the propane-3-yl
attachment
[0140] Among other things, the structure of the UNA monomer allows
it to be attached to naturally-occurring nucleotides.
[0141] A UNA oligomer can be a chain composed of UNA monomers, as
well as various nucleotides that may be based on
naturally-occurring nucleosides.
[0142] In some embodiments, the functional group R.sup.3 of a UNA
monomer can be --OR.sup.4, --SR.sup.4, --NR.sup.4.sub.2,
--NH(C.dbd.O)R.sup.4, morpholino, morpholin-1-yl, piperazin-1-yl,
or 4-alkanoyl-piperazin-1-yl, where R.sup.4 is the same or
different for each occurrence, and can be H, alkyl, a cholesterol,
a lipid molecule, a polyamine, an amino acid, or a polypeptide.
[0143] The UNA monomers are organic molecules. UNA monomers are not
nucleic acid monomers or nucleotides, nor are they
naturally-occurring nucleosides or modified naturally-occurring
nucleosides.
[0144] A UNA oligomer of this invention is a synthetic chain
molecule.
[0145] In some embodiments, as shown above, a UNA monomer can be
UNA-A (designated ), UNA-U (designated ), UNA-C (designated {hacek
over (C)}) and UNA-G (designated {hacek over (G)}).
[0146] Designations that may be used herein include mA, mG, mC, and
mU, which refer to the 2'-O-Methyl modified ribonucleotides.
[0147] Designations that may be used herein include dT, which
refers to a 2'-deoxy T nucleotide.
[0148] Additional Monomers for Oligomers
[0149] As used herein, in the context of oligomer sequences, the
symbol X represents a UNA monomer. When a mUNA oligomer is
complexed or duplexed with a nucleic acid molecule, the UNA
monomers of the mUNA oligomer can have any base attached that would
be complementary to the monomer with which it is paired in the
nucleic acid molecule.
[0150] As used herein, in the context of oligomer sequences, the
symbol N can represent any natural nucleotide monomer, or any
modified nucleotide monomer. When a mUNA oligomer is complexed or
duplexed with a nucleic acid molecule, an N monomer of the mUNA
oligomer can have any base attached that would be complementary to
the monomer with which it is paired in the nucleic acid
molecule.
[0151] As used herein, in the context of oligomer sequences, the
symbol Q represents a non-natural, modified, or chemically-modified
nucleotide monomer. When a mUNA oligomer is complexed or duplexed
with a nucleic acid molecule, a Q monomer of the mUNA oligomer can
have any base attached that would be complementary to the monomer
with which it is paired in the nucleic acid molecule.
[0152] Examples of nucleic acid monomers include non-natural,
modified, and chemically-modified nucleotides, including any such
nucleotides known in the art.
[0153] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include any such nucleotides known in the art,
for example, 2'-O-methyl ribonucleotides, 2'-O-methyl purine
nucleotides, 2'-deoxy-2'-fluoro ribonucleotides, 2'-deoxy-2'-fluoro
pyrimidine nucleotides, 2'-deoxy ribonucleotides, 2'-deoxy purine
nucleotides, universal base nucleotides, 5-C-methyl-nucleotides,
and inverted deoxyabasic monomer residues.
[0154] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include 3'-end stabilized nucleotides,
3'-glyceryl nucleotides, 3'-inverted abasic nucleotides, and
3'-inverted thymidine.
[0155] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include locked nucleic acid nucleotides (LNA),
2'-O,4'-C-methylene-(D-ribofuranosyl) nucleotides, 2'-methoxyethoxy
(MOE) nucleotides, 2'-methyl-thio-ethyl, 2'-deoxy-2'-fluoro
nucleotides, and 2'-O-methyl nucleotides.
[0156] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include 2', 4'-Constrained 2'-O-Methoxyethyl
(cMOE) and 2'-O-Ethyl (cEt) Modified DNAs.
[0157] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include 2'-amino nucleotides, 2'-O-amino
nucleotides, 2'-C-allyl nucleotides, and 2'-O-allyl
nucleotides.
[0158] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include N.sup.6-methyladenosine
nucleotides.
[0159] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include nucleotide monomers with modified bases
5-(3-amino)propyluridine, 5-(2-mercapto)ethyluridine,
5-bromouridine; 8-bromoguanosine, or 7-deazaadenosine.
[0160] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include 2'-O-aminopropyl substituted
nucleotides.
[0161] Examples of non-natural, modified, and chemically-modified
nucleotide monomers include replacing the 2'-OH group of a
nucleotide with a 2'-R, a 2'-OR, a 2'-halogen, a 2'-SR, or a
2'-amino, where R can be H, alkyl, alkenyl, or alkynyl.
[0162] Examples of nucleotide monomers include pseudouridine
(psi-Uridine) and 1-methylpseudouridine.
[0163] Some examples of modified nucleotides are given in Saenger,
Principles of Nucleic Acid Structure, Springer-Verlag, 1984.
[0164] mUNA Compounds
[0165] Aspects of this invention provide structures and
compositions for mUNA molecules that are oligomeric compounds. The
mUNA compounds can be active agents for pharmaceutical
compositions.
[0166] An oligomeric mUNA agent of this invention may contain one
or more UNA monomers. Oligomeric molecules of this invention can be
used as active agents in formulations for supplying peptide and
protein therapeutics.
[0167] In some embodiments, this invention provides oligomeric mUNA
compounds having a structure that incorporates novel combinations
of UNA monomers with certain natural nucleotides, non-natural
nucleotides, modified nucleotides, or chemically-modified
nucleotides.
[0168] Oligomeric mUNA compounds of this invention can have a
length of from about 200 to about 12,000 bases in length.
Oligomeric mUNA compounds of this invention can have a length of
about 1800, or about 1900, or about 2000, or about 2100, or about
2200, or about 2300, or about 2400, or about 2500 bases.
[0169] In further aspects, the oligomeric mUNA compounds of this
invention can be pharmacologically active molecules. A mUNA
molecule can be used as an active pharmaceutical ingredient for
generating a peptide or protein active agent in vitro, in vivo, or
ex vivo.
[0170] A mUNA molecule of this invention can have the structure of
Formula I
##STR00006##
wherein L.sup.1 is a linkage, n is from 200 to 12,000, and for each
occurrence L.sup.2 is a UNA linker group having the formula
--C.sup.1--C.sup.2--C.sup.3--, where R is attached to C.sup.2 and
has the formula --OCH(CH.sub.2R.sup.3)R.sup.5, where R.sup.3 is
--OR.sup.4, --SR.sup.4, --NR.sup.4.sub.2, --NH(C.dbd.O)R.sup.4,
morpholino, morpholin-1-yl, piperazin-1-yl, or
4-alkanoyl-piperazin-1-yl, where R.sup.4 is the same or different
for each occurrence and is H, alkyl, a cholesterol, a lipid
molecule, a polyamine, an amino acid, or a polypeptide, and where
R.sup.5 is a nucleobase, or L.sup.2(R) is a sugar such as a ribose
and R is a nucleobase, or L.sup.2 is a modified sugar such as a
modified ribose and R is a nucleobase. In certain embodiments, a
nucleobase can be a modified nucleobase. L.sup.1 can be a
phosphodiester linkage.
[0171] The base sequence of a mUNA molecule can be any sequence of
nucleobases.
[0172] In some aspects, a mUNA molecule of this invention can have
any number of phosphorothioate intermonomer linkages in any
intermonomer location.
[0173] In some embodiments, any one or more of the intermonomer
linkages of a mUNA molecule can be a phosphodiester, a
phosphorothioate including dithioates, a chiral phosphorothioate,
and other chemically modified forms.
[0174] When a mUNA molecule terminates in a UNA monomer, the
terminal position has a 1-end, or the terminal position has a
3-end, according to the positional numbering shown above.
[0175] mUNA Molecules with Enhanced Translation
[0176] A mUNA molecule of this invention can incorporate a region
that enhances the translational efficiency of the mUNA
molecule.
[0177] In general, translational enhancer regions as known in the
art can be incorporated into the structure of a mUNA molecule to
increase peptide or protein yields.
[0178] A mUNA molecule containing a translation enhancer region can
provide increased production of peptide or protein.
[0179] In some embodiments, a translation enhancer region can
comprise, or be located in a 5' or 3' untranslated region of a mUNA
molecule.
[0180] Examples of translation enhancer regions include
naturally-occurring enhancer regions from TEV 5'UTR and Xenopus
beta-globin 3'UTR.
[0181] mUNA Molecular Structure and Sequences
[0182] A mUNA molecule can be designed to express a target peptide
or protein. In some embodiments, the target peptide or protein can
be associated with a condition or disease in a subject.
[0183] In some aspects, the base sequence of a mUNA molecule can
include a portion that is identical to at least an effective
portion or domain of a base sequence of an mRNA, where an effective
portion is sufficient to impart a therapeutic activity to a
translation product of the mUNA molecule.
[0184] In some aspects, this invention provides active mUNA
oligomer molecules having a base sequence identical to at least a
fragment of a native nucleic acid molecule of a cell.
[0185] In certain embodiments, the base sequence of a mUNA molecule
can include a portion that is identical to a base sequence of an
mRNA, except for one or more base mutations. The number of
mutations for the mUNA molecule should not exceed an amount that
would produce a translation product of the mUNA molecule having
substantially less activity than the mRNA.
[0186] The oligomer mUNA molecules of this invention can display a
sequence of nucleobases, and can be designed to express a peptide
or protein, in vitro, ex vivo, or in vivo. The expressed peptide or
protein can have activity in various forms, including activity
corresponding to protein expressed from a native or natural
mRNA.
[0187] In some embodiments, a mUNA molecule of this invention may
have a chain length of about 400 to 15,000 monomers, where any
monomer that is not a UNA monomer can be a Q monomer.
[0188] mUNA Molecular Cap Structure
[0189] A mUNA molecule of this invention may have a 5'-end capped
with various groups and their analogues as are known in the art.
The 5' cap may be a m7GpppGm cap. The 5' cap may be an ARCA cap
(3'-OMe-m7G(5')pppG). The 5' cap may be an mCAP (m7G(5')ppp(5')G,
N.sup.7-Methyl-Guanosine-5'-Triphosphate-5'-Guanosine). The 5' cap
may be resistant to hydrolysis.
[0190] Some examples of 5' cap structures are given in
WO2015/051169A2.
[0191] Genetic Basis for mUNA Molecules
[0192] In some embodiments, the mUNA molecules of this invention
can be structured to provide peptides or proteins that are
nominally expressed by any portion of a genome. Examples of genes
for which a mUNA molecule can be used to express the corresponding
peptide or protein are set forth below.
[0193] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Neoplasia,
PTEN; ATM; ATR; EGFR; ERBB2; ERBB3; ERBB4; Notch1; Notch2; Notch3;
Notch4; AKT; AKT2; AKT3; HIF; HIF1a; HIF3a; Met; HRG; Bc12; PPAR
alpha; PPAR gamma; WT1 (Wilms Tumor); FGF Receptor Family members
(5 members: 1, 2, 3, 4, 5); CDKN2a; APC; RB (retinoblastoma); MEN1;
VHL; BRCA1; BRCA2; AR (Androgen Receptor); TSG101; IGF; IGF
Receptor; Igf1 (4 variants); Igf2 (3 variants); Igf 1 Receptor; Igf
2 Receptor; Bax; Bc12; caspases family (9 members: 1, 2, 3, 4, 6,
7, 8, 9, 12); Kras; Apc.
[0194] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Age-related
Macular Degeneration, Schizophrenia, Aber; Cc12; Cc2; cp
(ceruloplasmin); Timp3; cathepsinD; Vldlr; Ccr2 Neuregulin1 (Nrg
1); Erb4 (receptor for Neuregulin); Complexin1 (Cplx1); Tph1
Tryptophan hydroxylase; Tph2 Tryptophan hydroxylase 2; Neurexin 1;
GSK3; GSK3a; GSK3b.
[0195] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: 5-HTT
(Slc6a4); COMT; DRD (Drd1a); SLC6A3; DAOA; DTNBP1; Dao (Dao1).
[0196] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Trinucleotide
Repeat Disorders, HTT (Huntington's Dx); SBMA/SMAX1/AR (Kennedy's
Dx); FXN/X25 (Friedrich's Ataxia); ATX3 (Machado-Joseph's Dx);
ATXN1 and ATXN2 (spinocerebellar ataxias); DMPK (myotonic
dystrophy); Atrophin-1 and Atn 1 (DRPLA Dx); CBP (Creb-BP-global
instability); VLDLR (Alzheimer's); Atxn7; Atxn10.
[0197] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Fragile X
Syndrome, FMR2; FXR1; FXR2; mGLUR5.
[0198] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Secretase
Related Disorders, APH-1 (alpha and beta); Presenilin (Psen1);
nicastrin (Ncstn); PEN-2.
[0199] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Nos1.
[0200] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Parp1.
[0201] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Nat1;
Nat2.
[0202] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Prion-related
disorders, Prp.
[0203] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: ALS disease,
SOD1; ALS2; STEX; FUS; TARDBP; VEGF (VEGF-a; VEGF-b; VEGF-c).
[0204] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Drug
addiction, Prkce (alcohol); Drd2; Drd4; ABAT (alcohol); GRIA2;
Grm5; Grin1; Htr1b; Grin2a; Drd3; Pdyn; Gria1 (alcohol).
[0205] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Autism,
Mecp2; BZRAP1; MDGA2; Sema5A; Neurexin 1; Fragile X (FMR2 (AFF2);
FXR1; FXR2; Mglur5).
[0206] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Alzheimer's
Disease, E1; CHIP; UCH; UBB; Tau; LRP; PICALM; Clusterin; PS1;
SORL1; CR1; Vld1r; Uba1; Uba3; CHIP28 (Aqp1, Aquaporin 1); Uch11;
Uch13; APP.
[0207] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Inflammation,
1L-10; IL-1 (1L-1a; IL-1b); 1L-13; IL-17 (IL-17a (CTLA8); IL-17b;
IL-17c; IL-17d; IL-17f); II-23; Cx3er1; ptpn22; TNFa; NOD2/CARD15
for IBD; IL-6; 1L-12 (1L-12a; 1L-12b); CTLA4; Cx3cl1.
[0208] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Parkinson's
Disease, x-Synuclein; DJ-1; LRRK2; Parkin; PINK1.
[0209] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Blood and
coagulation diseases and disorders, Anemia (CDAN1, CDA1, RPS19,
DBA, PKLR, PK1, NT5C3, UMPH1, PSN1, RHAG, RH50A, NRAMP2, SPTB,
ALAS2, ANH1, ASB, ABCB7, ABC7, ASAT); Bare lymphocyte syndrome
(TAPBP, TPSN, TAP2, ABCB3, PSF2, RING11, MHC2TA, C2TA, RFX5, RFXAP,
RFX5), Bleeding disorders (TBXA2R, P2RX1, P2X1); Factor H and
factor H-like 1 (HF1, CFH, HUS); Factor V and factor VIII (MCFD2);
Factor VII deficiency (F7); Factor X deficiency (F10); Factor XI
deficiency (F11); Factor XII deficiency (F12, HAF); Factor XIIIA
deficiency (F13A1, F13A); Factor XIIIB deficiency (F13B); Fanconi
anemia (FANCA, FACA, FA1, FA, FAA, FAAP95, FAAP90, FLJ34064, FANCB,
FANCC, FACC, BRCA2, FANCD1, FANCD2, FANCD, FACD, FAD, FANCE, FACE,
FANCF, XRCC9, FANCG, BRIP1, BACH1, FANCJ, PHF9, FANCL, FANCM,
KIAA1596); Hemophagocytic lymphohistiocytosis disorders (PRF1,
HPLH2, UNC13D, MUNC13-4, HPLH3, HLH3, FHL3); Hemophilia A (F8, F8C,
HEMA); Hemophilia B (F9 Factor IX, HEMB), Hemorrhagic disorders
(PI, ATT, F5); Leukocyde deficiencies and disorders (ITGB2, CD18,
LCAMB, LAD, EIF2B1, EIF2BA, EIF2B2, EIF2B3, EIF2B5, LVWM, CACH,
CLE, EIF2B4); Sickle cell anemia (HBB); Thalassemia (HBA2, HBB,
HBD, LCRB, HBA1).
[0210] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Cell
dysregulation and oncology diseases and disorders, B-cell
non-Hodgkin lymphoma (BCL7A, BCL7); Leukemia (TAL1 TCL5, SCL, TAL2,
FLT3, NBS1, NBS, ZNFN1A1, IK1, LYF1, HOXD4, HOX4B, BCR, CML, PHL,
ALL, ARNT, KRAS2, RASK2, GMPS, AF10, ARHGEF12, LARG, KIAA0382,
CALM, CLTH, CEBPA, CEBP, CHIC2, BTL, FLT3, KIT, PBT, LPP, NPM1,
NUP214, D9S46E, CAN, CAIN, RUNX1, CBFA2, AML1, WHSC1L1, NSD3, FLT3,
AF1Q, NPM1, NUMA1, ZNF145, PLZF, PML, MYL, STAT5B, AF10, CALM,
CLTH, ARL11, ARLTS1, P2RX7, P2X7, BCR, CML, PHL, ALL, GRAF, NF1,
VRNF, WSS, NFNS, PTPN11, PTP2C, SHP2, NS1, BCL2, CCND1, PRAD1,
BCL1, TCRA, GATA1, GF1, ERYF1, NFE1, ABL1, NQO1, DIA4, NMOR1,
NUP214, D9S46E, CAN, CAIN).
[0211] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Inflammation
and immune related diseases and disorders, AIDS (KIR3DL1, NKAT3,
NKB1, AMB11, KIR3DS1, IFNG, CXCL12, SDF1); Autoimmune
lymphoproliferative syndrome (TNFRSF6, APT1, FAS, CD95, ALPS1A);
Combined immuno-deficiency, (IL2RG, SCIDX1, SCIDX, IMD4); HIV-1
(CCL5, SCYA5, D17S136E, TCP228), HIV susceptibility or infection
(IL10, CSIF, CMKBR2, CCR2, CMKBR5, CCCKR5 (CCR5));
Immuno-deficiencies (CD3E, CD3G, AICDA, AID, HIGM2, TNFRSF5, CD40,
UNG, DGU, HIGM4, TNFSF5, CD40LG, HIGM1, IGM, FOXP3, IPEX, AIID,
XPID, PIDX, TNFRSF14B, TACI); Inflammation (IL-10, IL-1 (IL-1a,
IL-1b), IL-13, IL-17 (IL-17a (CTLA8), IL-17b, IL-17c, IL-17d,
IL-17f, II-23, Cx3cr1, ptpn22, TNFa, NOD2/CARD15 for IBD, IL-6,
IL-12 (IL-12a, IL-12b), CTLA4, Cx3c11); Severe combined
immunodeficiencies (SCIDs) (JAK3, JAKL, DCLRE1C, ARTEMIS, SCIDA,
RAG1, RAG2, ADA, PTPRC, CD45, LCA, IL7R, CD3D, T3D, IL2RG, SCIDX1,
SCIDX, IMD4).
[0212] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Metabolic,
liver, kidney and protein diseases and disorders, Amyloid
neuropathy (TTR, PALB); Amyloidosis (APOA1, APP, AAA, CVAP, AD1,
GSN, FGA, LYZ, TTR, PALB); Cirrhosis (KRT18, KRT8, CIRH1A, NAIC,
TEX292, KIAA1988); Cystic fibrosis (CFTR, BG213071, ABCC7, CF,
MRP7); Glycogen storage diseases (SLC2A2, GLUT2, G6PC, G6PT, G6PT1,
GAA, LAMP2, LAMPB, AGL, GDE, GBE1, GYS2, PYGL, PFKM); Hepatic
adenoma, 142330 (TCF1, HNF1A, MODY3), Hepatic failure, early onset,
and neurologic disorder (SCOD1, SCO1), Hepatic lipase deficiency
(LIPC), Hepato-blastoma, cancer and carcinomas (CTNNB1, PDGFRL,
PDGRL, PRLTS, AXIN1, AXIN, CTNNB1, TP53, P53, LFS1, IGF2R, MPRI,
MET, CASP8, MCH5; Medullary cystic kidney disease (UMOD, HNFJ,
FJHN, MCKD2, ADMCKD2); Phenylketonuria (PAH, PKU1, QDPR, DHPR,
PTS); Polycystic kidney and hepatic disease (FCYT, PKHD1, ARPKD,
PKD1, PKD2, PKD4, PKDTS, PRKCSH, G19P1, PCLD, SEC63).
[0213] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Lipoprotein
lipase, APOA1, APOC3 and APOA4.
[0214] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Muscular/skeletal diseases and disorders, Becker muscular dystrophy
(DMD, BMD, MYF6), Duchenne Muscular Dystrophy (DMD, BMD);
Emery-Dreifuss muscular dystrophy (LMNA, LMN1, EMD2, FPLD, CMD1A,
HGPS, LGMD1B, LMNA, LMN1, EMD2, FPLD, CMD1A); Facio-scapulohumeral
muscular dystrophy (FSHMD1A, FSHD1A); Muscular dystrophy (FKRP,
MDC1C, LGMD2I, LAMA2, LAMM, LARGE, KIAA0609, MDC1D, FCMD, TTID,
MYOT, CAPN3, CANP3, DYSF, LGMD2B, SGCG, LGMD2C, DMDA1, SCG3, SGCA,
ADL, DAG2, LGMD2D, DMDA2, SGCB, LGMD2E, SGCD, SGD, LGMD2F, CMD1L,
TCAP, LGMD2G, CMD1N, TRIM32, HT2A, LGMD2H, FKRP, MDC1C, LGMD2I,
TTN, CMD1G, TMD, LGMD2J, POMT1, CAV3, LGMD1C, SEPN1, SELN, RSMD1,
PLEC1, PLTN, EBS1); Osteopetrosis (LRP5, BMND1, LRP7, LR3, OPPG,
VBCH2, CLCN7, CLC7, OPTA2, OSTM1, GL, TCIRG1, TIRC7, OC116, OPTB1);
Muscular atrophy (VAPB, VAPC, ALS8, SMN1, SMA1, SMA2, SMA3, SMA4,
BSCL2, SPG17, GARS, SMAD1, CMT2D, HEXB, IGHMBP2, SMUBP2, CATF1,
SMARD1).
[0215] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Neurological
and neuronal diseases and disorders, ALS (SOD1, ALS2, STEX, FUS,
TARDBP, VEGF (VEGF-a, VEGF-b, VEGF-c); Alzheimer's Disease (APP,
AAA, CVAP, AD1, APOE, AD2, PSEN2, AD4, STM2, APBB2, FE65L1, NOS3,
PLAU, URK, ACE, DCP1, ACE1, MPO, PACIP1, PAXIP1L, PTIP, A2M, BLMH,
BMH, PSEN1, AD3); Autism (Mecp2, BZRAP1, MDGA2, Sema5A, Neurexin 1,
GLO1, MECP2, RTT, PPMX, MRX16, MRX79, NLGN3, NLGN4, KIAA1260,
AUTSX2); Fragile X Syndrome (FMR2, FXR1, FXR2, mGLUR5);
Huntington's disease and disease like disorders (HD, IT15, PRNP,
PRIP, JPH3, JP3, HDL2, TBP, SCA17); Parkinson disease (NR4A2,
NURR1, NOT, TINUR, SNCAIP, TBP, SCA17, SNCA, NACP, PARK1, PARK4,
DJ1, PARK7, LRRK2, PARK8, PINK1, PARK6, UCHL1, PARK5, SNCA, NACP,
PARK1, PARK4, PRKN, PARK2, PDJ, DBH, NDUFV2); Rett syndrome (MECP2,
RTT, PPMX, MRX16, MRX79, CDKL5, STK9, MECP2, RTT, PPMX, MRX16,
MRX79, x-Synuclein, DJ-1); Schizo-phrenia (Neuregulin1 (Nrg1), Erb4
(receptor for Neuregulin), Complexin1 (Cplx1), Tph1 Trypto-phan
hydroxylase, Tph2, Tryptophan hydroxylase 2, Neurexin 1, GSK3,
GSK3a, GSK3b, 5-HTT (S1c6a4), COMT, DRD (Drd1a), SLC6A3, DAOA,
DTNBP1, Dao (Dao1)); Secretase Related Dis-orders (APH-1 (alpha and
beta), Presenilin (Psen1), nicastrin, (Ncstn), PEN-2, Nos1, Parp1,
Nat1, Nat2); Trinucleotide Repeat Disorders (HTT (Huntington's Dx),
SBMA/SMAX1/AR (Kennedy's Dx), FXN/X25 (Friedrich's Ataxia), ATX3
(Machado-Joseph's Dx), ATXN1 and ATXN2 (spinocerebellar ataxias),
DMPK (myotonic dystrophy), Atrophin-1 and Atn1 (DRPLA Dx), CBP
(Creb-BP--global instability), VLDLR (Alzheimer's), Atxn7,
Atxn10).
[0216] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Occular
diseases and disorders, Age-related macular degeneration (Aber,
Cc12, Cc2, cp (ceruloplasmin), Timp3, cathepsinD, Vldlr, Ccr2);
Cataract (CRYAA, CRYA1, CRYBB2, CRYB2, PITX3, BFSP2, CP49, CP47,
CRYAA, CRYA1, PAX6, AN2, MGDA, CRYBA1, CRYB1, CRYGC, CRYG3, CCL,
LIM2, MP19, CRYGD, CRYG4, BFSP2, CP49, CP47, HSF4, CTM, HSF4, CTM,
MIP, AQP0, CRYAB, CRYA2, CTPP2, CRYBB1, CRYGD, CRYG4, CRYBB2,
CRYB2, CRYGC, CRYG3, CCL, CRYAA, CRYA1, GJA8, CX50, CAE1, GJA3,
CX46, CZP3, CAE3, CCM1, CAM, KRIT1); Corneal clouding and dystrophy
(APOA1, TGFBI, CSD2, CDGG1, CSD, BIGH3, CDG2, TACSTD2, TROP2, M1S1,
VSX1, RINX, PPCD, PPD, KTCN, COL8A2, FECD, PPCD2, PIP5K3, CFD);
Cornea plana congenital (KERA, CNA2); Glaucoma (MYOC, TIGR, GLC1A,
JOAG, GPOA, OPTN, GLC1E, FIP2, HYPL, NRP, CYP1B1, GLC3A, OPA1, NTG,
NPG, CYP1B1, GLC3A); Leber congenital amaurosis (CRB1, RP12, CRX,
CORD2, CRD, RPGRIP1, LCA6, CORD9, RPE65, RP20, AIPL1, LCA4, GUCY2D,
GUC2D, LCA1, CORD6, RDH12, LCA3); Macular dystrophy (ELOVL4, ADMD,
STGD2, STGD3, RDS, RP7, PRPH2, PRPH, AVMD, AOFMD, VMD2).
[0217] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Epilepsy,
myoclonic, EPM2A, MELF, EPM2 Lafora type, 254780 Epilepsy,
myoclonic, NHLRC1, EPM2A, EPM2B Lafora type, 254780.
[0218] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Duchenne
muscular DMD, BMD dystrophy, 310200 (3) AIDS, delayed/rapid
KIR3DL1, NKAT3, NKB1, AMB11, KIR3DS1 progression to (3).
[0219] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: AIDS,
delayed/rapid KIR3DL1, NKAT3, NKB1, AMB11, KIR3DS1 progression to
(3) AIDS, rapid IFNG progression to, 609423 (3) AIDS, resistance to
CXCL12, SDF1 (3).
[0220] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Alpha-1-Antitrypsin Deficiency, SERPINA1 [serpin peptidase
inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member
1]; SERPINA2 [serpin peptidase inhibitor, clade A (alpha-1
antiproteinase, antitrypsin), member 2]; SERPINA3 [serpin peptidase
inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member
3]; SERPINA5 [serpin peptidase inhibitor, clade A (alpha-1
antiproteinase, antitrypsin), member 5]; SERPINA6 [serpin peptidase
inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member
6]; SERPINA7 [serpin peptidase inhibitor, clade A (alpha-1
antiproteinase, antitrypsin), member 7];" AND "SERPLNA6 (serpin
peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin),
member 6).
[0221] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: PI3K/AKT
Signaling, PRKCE; ITGAM; ITGA5; IRAK1; PRKAA2; EIF2AK2; PTEN;
EIF4E; PRKCZ; GRK6; MAPK1; TSC1; PLK1; AKT2; IKBKB; PIK3CA; CDK8;
CDKN1B; NFKB2; BCL2; PIK3CB; PPP2R1A; MAPK8; BCL2L1; MAPK3; TSC2;
ITGA1; KRAS; EIF4EBP1; RELA; PRKCD; NOS3; PRKAA1; MAPK9; CDK2;
PPP2CA; PIM1; ITGB7; YWHAZ; ILK; TP53; RAF1.; IKBKG; RELB; DYRK1A;
CDKN1A; ITGB1; MAP2K2; JAK1; AKT1; JAK2; PIK3R1; CHUK; PDPK1;
PPP2R5C; CTNNB1.; MAP2K1; NFKB1; PAK3; ITGB3; CCND1; GSK3A; FRAP1;
SFN; ITGA2; TTK; CSNK1A1; BRAF; GSK3B; AKT3; FOXO1; SGK; HSP90AA1;
RPS6KB1.
[0222] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: ERK/MAPK
Signaling, PRKCE; ITGAM; ITGA5; HSPB1; IRAK1; PRKAA2; EIF2AK2;
RAC1; RAP1A; TLN1; EIF4E; ELK1; GRK6; MAPK1; RAC2; PLK1; AKT2;
PIK3CA; CDK8; CREB1; PRKCI; PTK2; FOS; RPS6KA4; PIK3CB; PPP2R1A;
PIK3C3; MAPK8; MAPK3; ITGA1; ETS1; KRAS; MYCN; EIF4EBP1; PPARG;
PRKCD; PRKAA1; MAPK9; SRC; CDK2; PPP2CA; PIM1; PIK3C2A; ITGB7;
YWHAZ; PPP1CC; KSR1; PXN; RAF1; FYN; DYRK1A; ITGB1; MAP2K2; PAK4;
PIK3R1; STAT3; PPP2R5C; MAP2K1; PAK3; ITGB3; ESR1; ITGA2; MYC; TTK;
CSNK1A1; CRKL; BRAF; ATF4; PRKCA; SRF; STAT1; SGK.
[0223] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Serine/Threonine-Protein Kinase, CDK16; PCTK1; CDK5R1.
[0224] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Glucocorticoid Receptor Signaling, RAC1; TAF4B; EP300; SMAD2;
TRAF6; PCAF; ELK1; MAPK1; SMAD3; AKT2; IKBKB; NCOR2; UBE2I; PIK3CA;
CREB1; FOS; HSPA5; NFKB2; BCL2; MAP3K14; STAT5B; PIK3CB; PIK3C3;
MAPK8; BCL2L1; MAPK3; TSC22D3; MAPK10; NRIP1; KRAS; MAPK13; RELA;
STAT5A; MAPK9; NOS2A; PBX1; NR3C1; PIK3C2A; CDKN1C; TRAF2;
SERPINE1; NCOA3; MAPK14; TNF; RAF1; IKBKG; MAP3K7; CREBBP; CDKN1A;
MAP2K2; JAK1; IL8; NCOA2; AKT1; JAK2; PIK3R1; CHUK; STAT3; MAP2K1;
NFKB1; TGFBR1; ESR1; SMAD4; CEBPB; JUN; AR; AKT3; CCL2; MMP1;
STAT1; IL6; HSP90AA1.
[0225] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Axonal
Guidance Signaling, PRKCE; ITGAM; ROCK1; ITGA5; CXCR4; ADAM12;
IGF1; RAC1; RAP1A; E1F4E; PRKCZ; NRP1; NTRK2; ARHGEF7; SMO; ROCK2;
MAPK1; PGF; RAC2; PTPN11; GNAS; AKT2; PIK3CA; ERBB2; PRKC1; PTK2;
CFL1; GNAQ; PIK3CB; CXCL12; PIK3C3; WNT11; PRKD1; GNB2L1; ABL1;
MAPK3; ITGA1; KRAS; RHOA; PRKCD; PIK3C2A; ITGB7; GLI2; PXN; VASP;
RAF1; FYN; ITGB1; MAP2K2; PAK4; ADAM17; AKT1; PIK3R1; GLI1; WNT5A;
ADAM10; MAP2K1; PAK3; ITGB3; CDC42; VEGFA; ITGA2; EPHA8; CRKL;
RND1; GSK3B; AKT3; PRKCA.
[0226] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Ephrin
Receptor Signaling, PRKCE; ITGAM; ROCK1; ITGA5; CXCR4; IRAK1;
PRKAA2; EIF2AK2; RAC1; RAP1A; GRK6; ROCK2; MAPK1; PGF; RAC2;
PTPN11; GNAS; PLK1; AKT2; DOK1; CDK8; CREB1; PTK2; CFL1; GNAQ;
MAP3K14; CXCL12; MAPK8; GNB2L1; ABL1; MAPK3; ITGA1; KRAS; RHOA;
PRKCD; PRKAA1; MAPK9; SRC; CDK2; PIM1; ITGB7; PXN; RAF1; FYN;
DYRK1A; ITGB1; MAP2K2; PAK4, AKT1; JAK2; STAT3; ADAM10; MAP2K1;
PAK3; ITGB3; CDC42; VEGFA; ITGA2; EPHA8; TTK; CSNK1A1; CRKL; BRAF;
PTPN13; ATF4; AKT3; SGK.
[0227] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Actin
Cytoskeleton Signaling, ACTN4; PRKCE; ITGAM; ROCK1; ITGA5; IRAK1;
PRKAA2; EIF2AK2; RAC1; INS; ARHGEF7; GRK6; ROCK2; MAPK1; RAC2;
PLK1; AKT2; PIK3CA; CDK8; PTK2; CFL1; PIK3CB; MYH9; DIAPH1; PIK3C3;
MAPK8; F2R; MAPK3; SLC9A1; ITGA1; KRAS; RHOA; PRKCD; PRKAA1; MAPK9;
CDK2; PIM1; PIK3C2A; ITGB7; PPP1CC; PXN; VIL2; RAF1; GSN; DYRK1A;
ITGB1; MAP2K2; PAK4; PIP5K1A; PIK3R1; MAP2K1; PAK3; ITGB3; CDC42;
APC; ITGA2; TTK; CSNK1A1; CRKL; BRAF; VAV3; SGK.
[0228] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Huntington's
Disease Signaling, PRKCE; IGF1; EP300; RCOR1.; PRKCZ; HDAC4; TGM2;
MAPK1; CAPNS1; AKT2; EGFR; NCOR2; SP1; CAPN2; PIK3CA; HDAC5; CREB1;
PRKC1; HSPA5; REST; GNAQ; PIK3CB; PIK3C3; MAPK8; IGF1R; PRKD1;
GNB2L1; BCL2L1; CAPN1; MAPK3; CASP8; HDAC2; HDAC7A; PRKCD; HDAC11;
MAPK9; HDAC9; PIK3C2A; HDAC3; TP53; CASP9; CREBBP; AKT1; PIK3R1;
PDPK1; CASP1; APAF1; FRAP1; CASP2; JUN; BAX; ATF4; AKT3; PRKCA;
CLTC; SGK; HDAC6; CASP3.
[0229] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Apoptosis
Signaling, PRKCE; ROCK1; BID; IRAK1; PRKAA2; EIF2AK2; BAK1; BIRC4;
GRK6; MAPK1; CAPNS1; PLK1; AKT2; IKBKB; CAPN2; CDK8; FAS; NFKB2;
BCL2; MAP3K14; MAPK8; BCL2L1; CAPN1; MAPK3; CASP8; KRAS; RELA;
PRKCD; PRKAA1; MAPK9; CDK2; PIM1; TP53; TNF; RAF1; IKBKG; RELB;
CASP9; DYRK1A; MAP2K2; CHUK; APAF1; MAP2K1; NFKB1; PAK3; LMNA;
CASP2; BIRC2; TTK; CSNK1A1; BRAF; BAX; PRKCA; SGK; CASP3; BIRC3;
PARP1.
[0230] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: B Cell
Receptor Signaling, RAC1; PTEN; LYN; ELK1; MAPK1; RAC2; PTPN11;
AKT2; IKBKB; PIK3CA; CREB1; SYK; NFKB2; CAMK2A; MAP3K14; PIK3CB;
PIK3C3; MAPK8; BCL2L1; ABL1; MAPK3; ETS1; KRAS; MAPK13; RELA;
PTPN6; MAPK9; EGR1; PIK3C2A; BTK; MAPK14; RAF1; IKBKG; RELB;
MAP3K7; MAP2K2; AKT1; PIK3R1; CHUK; MAP2K1; NFKB1; CDC42; GSK3A;
FRAP1; BCL6; BCL10; JUN; GSK3B; ATF4; AKT3; VAV3; RPS6KB1.
[0231] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Leukocyte
Extravasation Signaling, ACTN4; CD44; PRKCE; ITGAM; ROCK1; CXCR4;
CYBA; RAC1; RAP1A; PRKCZ; ROCK2; RAC2; PTPN11; MMP14; PIK3CA;
PRKCI; PTK2; PIK3CB; CXCL12; PIK3C3; MAPK8; PRKD1; ABL1; MAPK10;
CYBB; MAPK13; RHOA; PRKCD; MAPK9; SRC; PIK3C2A; BTK; MAPK14; NOX1;
PXN; VIL2; VASP; ITGB1; MAP2K2; CTNND1; PIK3R1; CTNNB1; CLDN1;
CDC42; F11R; ITK; CRKL; VAV3; CTTN; PRKCA; MMP1; MMP9.
[0232] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Integrin
Signaling, ACTN4; ITGAM; ROCK1; ITGA5; RAC1; PTEN; RAP1A; TLN1;
ARHGEF7; MAPK1; RAC2; CAPNS1; AKT2; CAPN2; P1K3CA; PTK2; PIK3CB;
PIK3C3; MAPK8; CAV1; CAPN1; ABL1; MAPK3; ITGA1; KRAS; RHOA; SRC;
PIK3C2A; ITGB7; PPP1CC; ILK; PXN; VASP; RAF1; FYN; ITGB1; MAP2K2;
PAK4; AKT1; PIK3R1; TNK2; MAP2K1; PAK3; ITGB3; CDC42; RND3; ITGA2;
CRKL; BRAF; GSK3B; AKT3.
[0233] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Acute Phase
Response Signaling, IRAK1; SOD2; MYD88; TRAF6; ELK1; MAPK1; PTPN11;
AKT2; IKBKB; PIK3CA; FOS; NFKB2; MAP3K14; PIK3CB; MAPK8; RIPK1;
MAPK3; IL6ST; KRAS; MAPK13; IL6R; RELA; SOCS1; MAPK9; FTL; NR3C1;
TRAF2; SERPINE1; MAPK14; TNF; RAF1; PDK1; IKBKG; RELB; MAP3K7;
MAP2K2; AKT1; JAK2; PIK3R1; CHUK; STAT3; MAP2K1; NFKB1; FRAP1;
CEBPB; JUN; AKT3; IL1R1; IL6.
[0234] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: PTEN
Signaling, ITGAM; ITGA5; RAC1; PTEN; PRKCZ; BCL2L11; MAPK1; RAC2;
AKT2; EGFR; IKBKB; CBL; PIK3CA; CDKN1B; PTK2; NFKB2; BCL2; PIK3CB;
BCL2L1; MAPK3; ITGA1; KRAS; ITGB7; ILK; PDGFRB; INSR; RAF1; IKBKG;
CASP9; CDKN1A; ITGB1; MAP2K2; AKT1; PIK3R1; CHUK; PDGFRA; PDPK1;
MAP2K1; NFKB1; ITGB3; CDC42; CCND1; GSK3A; ITGA2; GSK3B; AKT3;
FOXO1; CASP3; RPS6KB1.
[0235] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: p53
Signaling, PTEN; EP300; BBC3; PCAF; FASN; BRCA1; GADD45A; BIRC5;
AKT2; PIK3CA; CHEK1; TP53INP1; BCL2; PIK3CB; PIK3C3; MAPK8; THBS1;
ATR; BCL2L1; E2F1; PMAIP1; CHEK2; TNFRSF10B; TP73; RBI; HDAC9;
CDK2; PIK3C2A; MAPK14; TP53; LRDD; CDKN1A; HIPK2; AKT1; RIK3R1;
RRM2B; APAF1; CTNNB1; SIRT1; CCND1; PRKDC; ATM; SFN; CDKN2A; JUN;
SNAI2; GSK3B; BAX; AKT3.
[0236] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Aryl
Hydrocarbon Receptor Signaling, HSPB1; EP300; FASN; TGM2; RXRA;
MAPK1; NQO1; NCOR2; SP1; ARNT; CDKN1B; FOS; CHEK1; SMARCA4; NFKB2;
MAPK8; ALDH1A1; ATR; E2F1; MAPK3; NRIP1; CHEK2; RELA; TP73; GSTP1;
RB1; SRC; CDK2; AHR; NFE2L2; NCOA3; TP53; TNF; CDKN1A; NCOA2;
APAF1; NFKB1; CCND1; ATM; ESR1; CDKN2A; MYC; JUN; ESR2; BAX; IL6;
CYP1B1; HSP90AA1.
[0237] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Xenobiotic
Metabolism Signaling, PRKCE; EP300; PRKCZ; RXRA; MAPK1; NQO1;
NCOR2; PIK3CA; ARNT; PRKCI; NFKB2; CAMK2A; PIK3CB; PPP2R1A; PIK3C3;
MAPK8; PRKD1; ALDH1A1; MAPK3; NRIP1; KRAS; MAPK13; PRKCD; GSTP1;
MAPK9; NOS2A; ABCB1; AHR; PPP2CA; FTL; NFE2L2; PIK3C2A; PPARGC1A;
MAPK14; TNF; RAF1; CREBBP; MAP2K2; PIK3R1; PPP2R5C; MAP2K1; NFKB1;
KEAP1; PRKCA; EIF2AK3; IL6; CYP1B1; HSP90AA1.
[0238] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: SAPK/JNK
Signaling, PRKCE; IRAK1; PRKAA2; EIF2AK2; RAC1; ELK1; GRK6; MAPK1;
GADD45A; RAC2; PLK1; AKT2; PIK3CA; FADD; CDK8; PIK3CB; PIK3C3;
MAPK8; RIPK1; GNB2L1; IRS1; MAPK3; MAPK10; DAXX; KRAS; PRKCD;
PRKAA1; MAPK9; CDK2; PIM1; PIK3C2A; TRAF2; TP53; LCK; MAP3K7;
DYRK1A; MAP2K2; PIK3R1; MAP2K1; PAK3; CDC42; JUN; TTK; CSNK1A1;
CRKL; BRAF; SGK.
[0239] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: PPAr/RXR
Signaling, PRKAA2; EP300; INS; SMAD2; TRAF6; PPARA; FASN; RXRA;
MAPK1; SMAD3; GNAS; IKBKB; NCOR2; ABCA1; GNAQ; NFKB2; MAP3K14;
STAT5B; MAPK8; IRS1; MAPK3; KRAS; RELA; PRKAA1; PPARGC1A; NCOA3;
MAPK14; INSR; RAF1; IKBKG; RELB; MAP3K7; CREBBP; MAP2K2; JAK2;
CHUK; MAP2K1; NFKB1; TGFBR1; SMAD4; JUN; IL1R1; PRKCA; IL6;
HSP90AA1; ADIPOQ.
[0240] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: NF-KB
Signaling, IRAK1; EIF2AK2; EP300; INS; MYD8; PRKCZ: TRAF6; TBK1;
AKT2; EGFR; IKBKB; PIK3CA; BTRC; NFKB2; MAP3K14; PIK3CB; PIK3C3;
MAPK8; RIPK1; HDAC2; KRAS; RELA; PIK3C2A; TRAF2; TLR4: PDGFRB; TNF;
INSR; LCK; IKBKG; RELB; MAP3K7; CREBBP; AKT1; PIK3R1; CHUK; PDGFRA;
NFKB1; TLR2; BCL10; GSK3B; AKT3; TNFAIP3; IL1R1.
[0241] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Neuregulin
Signaling, ERBB4; PRKCE; ITGAM; ITGA5: PTEN; PRKCZ; ELK1; MAPK1;
PTPN11; AKT2; EGFR; ERBB2; PRKCI; CDKN1B; STAT5B; PRKD1; MAPK3;
ITGA1; KRAS; PRKCD; STAT5A; SRC; ITGB7; RAF1; ITGB1; MAP2K2;
ADAM17; AKT1; PIK3R1; PDPK1; MAP2K1; ITGB3; EREG; FRAP1; PSEN1;
ITGA2; MYC; NRG1; CRKL; AKT3; PRKCA; HSP90AA1; RPS6KB1.
[0242] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Wnt &
Beta catenin Signaling, CD44; EP300; LRP6; DVL3; CSNK1E; GJA1; SMO;
AKT2; PIN1; CDH1; BTRC; GNAQ; MARK2; PPP2R1A; WNT11; SRC; DKK1;
PPP2CA; SOX6; SFRP2: ILK; LEF1; SOX9; TP53; MAP3K7; CREBBP; TCF7L2;
AKT1; PPP2R5C; WNT5A; LRP5; CTNNB1; TGFBR1; CCND1; GSK3A; DVL1;
APC; CDKN2A; MYC; CSNK1A1; GSK3B; AKT3; SOX2.
[0243] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Insulin
Receptor Signaling, PTEN; INS; EIF4E; PTPN1; PRKCZ; MAPK1; TSC1;
PTPN11; AKT2; CBL; PIK3CA; PRKCI; PIK3CB; PIK3C3; MAPK8; IRS1;
MAPK3; TSC2; KRAS; EIF4EBP1; SLC2A4; PIK3C2A; PPP1CC; INSR; RAF1;
FYN; MAP2K2; JAK1; AKT1; JAK2; PIK3R1; PDPK1; MAP2K1; GSK3A; FRAP1;
CRKL; GSK3B; AKT3; FOXO1; SGK; RPS6KB1.
[0244] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: IL-6
Signaling, HSPB1; TRAF6; MAPKAPK2; ELK1; MAPK1; PTPN11; IKBKB; FOS;
NFKB2: MAP3K14; MAPK8; MAPK3; MAPK10; IL6ST; KRAS; MAPK13; IL6R;
RELA; SOCS1; MAPK9; ABCB1; TRAF2; MAPK14; TNF; RAF1; IKBKG; RELB;
MAP3K7; MAP2K2; IL8; JAK2; CHUK; STAT3; MAP2K1; NFKB1; CEBPB; JUN;
IL1R1; SRF; IL6.
[0245] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Hepatic
Cholestasis, PRKCE; IRAK1; INS; MYD8; PRKCZ; TRAF6; PPARA; RXRA;
IKBKB; PRKCI; NFKB2; MAP3K14; MAPK8; PRKD1; MAPK10; RELA; PRKCD;
MAPK9; ABCB1; TRAF2; TLR4; TNF; INSR; IKBKG; RELB; MAP3K7; IL8;
CHUK; NR1H2; TJP2; NFKB1; ESR1; SREBF1; FGFR4; JUN; IL1R1; PRKCA;
IL6.
[0246] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: IGF-1
Signaling, IGF1; PRKCZ; ELK1; MAPK1; PTPN11; NEDD4; AKT2; PIK3CA;
PRKC1; PTK2; FOS; PIK3CB; PIK3C3; MAPK8; 1GF1R; IRS1; MAPK3;
IGFBP7; KRAS; PIK3C2A; YWHAZ; PXN; RAF1; CASP9; MAP2K2; AKT1;
PIK3R1; PDPK1; MAP2K1; IGFBP2; SFN; JUN; CYR61; AKT3; FOXO1; SRF;
CTGF; RPS6KB1.
[0247] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: NRF2-mediated
Oxidative Stress Response, PRKCE; EP300; SOD2; PRKCZ; MAPK1;
SQSTM1; NQO1; PIK3CA; PRKC1; FOS; PIK3CB; P1K3C3; MAPK8; PRKD1;
MAPK3; KRAS; PRKCD; GSTP1; MAPK9; FTL; NFE2L2; PIK3C2A; MAPK14;
RAF1; MAP3K7; CREBBP; MAP2K2; AKT1; PIK3R1; MAP2K1; PPIB; JUN;
KEAP1; GSK3B; ATF4; PRKCA; EIF2AK3; HSP90AA1.
[0248] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Hepatic,
Fibrosis/Hepatic Stellate Cell Activation, EDN1; IGF1; KDR; FLT1;
SMAD2; FGFR1; MET; PGF; SMAD3; EGFR; FAS; CSF1; NFKB2; BCL2; MYH9;
IGF1R; IL6R; RELA; TLR4; PDGFRB; TNF; RELB; IL8; PDGFRA; NFKB1;
TGFBR1; SMAD4; VEGFA; BAX; IL1R1; CCL2; HGF; MMP1; STAT1; IL6;
CTGF; MMP9.
[0249] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: PPAR
Signaling, EP300; INS; TRAF6; PPARA; RXRA; MAPK1; IKBKB; NCOR2;
FOS; NFKB2; MAP3K14; STAT5B; MAPK3; NRIP1; KRAS; PPARG; RELA;
STAT5A; TRAF2; PPARGC1A; PDGFRB; TNF; INSR; RAF1; IKBKG; RELB;
MAP3K7; CREBBP; MAP2K2; CHUK; PDGFRA; MAP2K1; NFKB1; JUN; IL1R1;
HSP90AA1.
[0250] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Fc Epsilon RI
Signaling, PRKCE; RAC1; PRKCZ; LYN; MAPK1; RAC2; PTPN11; AKT2;
PIK3CA; SYK; PRKCI; PIK3CB; PIK3C3; MAPK8; PRKD1; MAPK3; MAPK10;
KRAS; MAPK13; PRKCD; MAPK9; PIK3C2A; BTK; MAPK14; TNF; RAF1; FYN;
MAP2K2; AKT1; PIK3R1; PDPK1; MAP2K1; AKT3; VAV3; PRKCA.
[0251] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: G-Protein
Coupled Receptor Signaling, PRKCE; RAP1A; RGS16; MAPK1; GNAS; AKT2;
IKBKB; PIK3CA; CREB1; GNAQ; NFKB2; CAMK2A; PIK3CB; PIK3C3; MAPK3;
KRAS; RELA; SRC; PIK3C2A; RAF1; IKBKG; RELB; FYN; MAP2K2; AKT1;
PIK3R1; CHUK; PDPK1; STAT3; MAP2K1; NFKB1; BRAF; ATF4; AKT3;
PRKCA.
[0252] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Inositol
Phosphate Metabolism, PRKCE; IRAK1; PRKAA2; EIF2AK2; PTEN; GRK6;
MAPK1; PLK1; AKT2; PIK3CA; CDK8; PIK3CB; PIK3C3; MAPK8; MAPK3;
PRKCD; PRKAA1; MAPK9; CDK2; PIM1; PIK3C2A; DYRK1A; MAP2K2; PIP5K1A;
PIK3R1; MAP2K1; PAK3; ATM; TTK; CSNK1A1; BRAF; SGK.
[0253] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: PDGF
Signaling, EIF2AK2; ELK1; ABL2; MAPK1; PIK3CA; FOS; PIK3CB;PIK3C3;
MAPK8; CAV1; ABL1; MAPK3; KRAS; SRC; PIK3C2A; PDGFRB; RAF1; MAP2K2;
JAK1; JAK2; PIK3R1; PDGFRA; STAT3; SPHK1; MAP2K1; MYC; JUN; CRKL;
PRKCA; SRF; STAT1; SPHK2.
[0254] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: VEGF
Signaling, ACTN4; ROCK1; KDR; FLT1; ROCK2; MAPK1; PGF; AKT2;
PIK3CA; ARNT; PTK2; BCL2; PIK3CB; PIK3C3; BCL2L1; MAPK3; KRAS;
HIF1A; NOS3; PIK3C2A; PXN; RAF1; MAP2K2; ELAVL1; AKT1; PIK3R1;
MAP2K1; SFN; VEGFA; AKT3; FOXO1; PRKCA.
[0255] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Natural
Killer Cell Signaling, PRKCE; RAC1; PRKCZ; MAPK1; RAC2; PTPN11;
KIR2DL3; AKT2; PIK3CA; SYK; PRKCI; PIK3CB; PIK3C3; PRKD1; MAPK3;
KRAS; PRKCD; PTPN6; PIK3C2A; LCK; RAF1; FYN; MAP2K2; PAK4; AKT1;
PIK3R1; MAP2K1; PAK3; AKT3; VAV3; PRKCA.
[0256] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Cell Cycle:
G1/S Checkpoint Regulation, HDAC4; SMAD3; SUV39H1; HDAC5; CDKN1B;
BTRC; ATR; ABL1; E2F1; HDAC2; HDAC7A; RB1; HDAC11; HDAC9; CDK2;
E2F2; HDAC3; TP53; CDKN1A; CCND1; E2F4; ATM; RBL2; SMAD4; CDKN2A;
MYC; NRG1; GSK3B; RBL1; HDAC6.
[0257] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: T Cell
Receptor Signaling, RAC1; ELK1; MAPK1; IKBKB; CBL; PIK3CA; FOS;
NFKB2; PIK3CB; PIK3C3; MAPK8; MAPK3; KRAS; RELA, PIK3C2A; BTK; LCK;
RAF1; IKBKG; RELB, FYN; MAP2K2; PIK3R1; CHUK; MAP2K1; NFKB1; ITK;
BCL10; JUN; VAV3.
[0258] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Death
Receptor Signaling, CRADD; HSPB1; BID; BIRC4; TBK1; IKBKB; FADD;
FAS; NFKB2; BCL2; MAP3K14; MAPK8; RIPK1; CASP8; DAXX; TNFRSF10B;
RELA; TRAF2; TNF; IKBKG; RELB; CASP9; CHUK; APAF1; NFKB1; CASP2;
BIRC2; CASP3; BIRC3.
[0259] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: FGF Signaling
RAC1; FGFR1; MET; MAPKAPK2; MAPK1; PTPN11; AKT2; PIK3CA; CREB1;
PIK3CB; PIK3C3; MAPK8; MAPK3; MAPK13; PTPN6; PIK3C2A; MAPK14; RAF1;
AKT1; PIK3R1; STAT3; MAP2K1; FGFR4; CRKL; ATF4; AKT3; PRKCA;
HGF.
[0260] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: GM-CSF
Signaling, LYN; ELK1; MAPK1; PTPN11; AKT2; PIK3CA; CAMK2A; STAT5B;
PIK3CB; PIK3C3; GNB2L1; BCL2L1; MAPK3; ETS1; KRAS; RUNX1; PIM1;
PIK3C2A; RAF1; MAP2K2; AKT1; JAK2; PIK3R1; STAT3; MAP2K1; CCND1;
AKT3; STAT1.
[0261] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Amyotrophic
Lateral Sclerosis Signaling, BID; IGF1; RAC1; BIRC4; PGF; CAPNS1;
CAPN2; PIK3CA; BCL2; PIK3CB; PIK3C3; BCL2L1; CAPN1; PIK3C2A; TP53;
CASP9; PIK3R1; RAB5A; CASP1; APAF1; VEGFA; BIRC2; BAX; AKT3; CASP3;
BIRC3.
[0262] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: JAK/Stat
Signaling, PTPN1; MAPK1; PTPN11; AKT2; PIK3CA; STAT5B; PIK3CB;
PIK3C3; MAPK3; KRAS; SOCS1; STAT5A; PTPN6; PIK3C2A; RAF1; CDKN1A;
MAP2K2; JAK1; AKT1; JAK2; PIK3R1; STAT3; MAP2K1; FRAP1; AKT3;
STAT1.
[0263] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Nicotinate
and Nicotinamide Metabolism, PRKCE; IRAK1; PRKAA2; EIF2AK2; GRK6;
MAPK1; PLK1; AKT2; CDK8; MAPK8; MAPK3; PRKCD; PRKAA1; PBEF1; MAPK9;
CDK2; PIM1; DYRK1A; MAP2K2; MAP2K1; PAK3; NT5E; TTK; CSNK1A1; BRAF;
SGK.
[0264] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Chemokine
Signaling, CXCR4; ROCK2; MAPK1; PTK2; FOS; CFL1; GNAQ; CAMK2A;
CXCL12; MAPK8; MAPK3; KRAS; MAPK13; RHOA; CCR3; SRC; PPP1CC;
MAPK14; NOXI; RAF1; MAP2K2; MAP2K1; JUN; CCL2; PRKCA.
[0265] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: IL-2
Signaling, ELK1; MAPK1; PTPN11; AKT2; PIK3CA; SYK; FOS; STAT5B;
PIK3CB; PIK3C3; MAPK8; MAPK3; KRAS; SOCS1; STAT5A; PIK3C2A: LCK;
RAF1; MAP2K2; JAK1; AKT1; PIK3R1; MAP2K1; JUN; AKT3.
[0266] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Synaptic Long
Term Depression, PRKCE; IGF1; PRKCZ; PRDX6; LYN; MAPK1; GNAS;
PRKC1; GNAQ; PPP2R1A; IGF1R; PRKID1; MAPK3; KRAS; GRN; PRKCD; NOS3;
NOS2A; PPP2CA; YWHAZ; RAF1; MAP2K2; PPP2R5C; MAP2K1; PRKCA.
[0267] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Estrogen
Receptor Signaling, TAF4B; EP300; CARM1; PCAF; MAPK1; NCOR2;
SMARCA4; MAPK3; NRIP1; KRAS; SRC; NR3C1; HDAC3; PPARGC1A; RBM9;
NCOA3; RAF1; CREBBP; MAP2K2; NCOA2; MAP2K1; PRKDC; ESR1; ESR2.
[0268] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Protein
Ubiquitination Pathway, TRAF6; SMURF1; BIRC4; BRCA1; UCHL1; NEDD4;
CBL; UBE2I; BTRC; HSPA5; USP7; USP10; FBXW7; USP9X; STUB1; USP22;
B2M; BIRC2; PARK2; USPS; USP1; VHL; HSP90AA1; BIRC3.
[0269] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: IL-10
Signaling, TRAF6; CCR1; ELK1; IKBKB; SP1; FOS; NFKB2; MAP3K14;
MAPK8; MAPK13; RELA; MAPK14; TNF; IKBKG; RELB; MAP3K7; JAK1; CHUK;
STAT3; NFKB1; JUN; IL1R1; IL6.
[0270] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: VDR/RXR
Activation, PRKCE; EP300; PRKCZ; RXRA; GADD45A; HEST; NCOR2; SP1;
PRKC1; CDKN1B; PRKD1; PRKCD; RUNX2; KLF4; YY1; NCOA3; CDKN1A;
NCOA2; SPP1; LRP5; CEBPB; FOXO1; PRKCA.
[0271] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: TGF-beta
Signaling, EP300; SMAD2; SMURF1; MAPK1; SMAD3; SMAD1; FOS; MAPK8;
MAPK3; KRAS; MAPK9; RUNX2; SERPINE1; RAF1; MAP3K7; CREBBP; MAP2K2;
MAP2K1; TGFBR1; SMAD4; JUN; SMAD5.
[0272] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Toll-like
Receptor Signaling, IRAK1; EIF2AK2; MYD8; TRAF6; PPARA; ELK1;
IKBKB; FOS; NFKB2; MAP3K14; MAPK8; MAPK13; RELA; TLR4; MAPK14;
IKBKG; RELB; MAP3K7; CHUK; NFKB1; TLR2; JUN.
[0273] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: p38 MAPK
Signaling, HSPB1; IRAK1; TRAF6; MAPKAPK2; ELK1; FADD; FAS; CREB1;
DDIT3; RPS6KA4; DAXX; MAPK13; TRAF2; MAPK14; TNF; MAP3K7; TGFBR1;
MYC; ATF4; IL1R1; SRF; STAT 1.
[0274] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Neurotrophin/TRK Signaling, NTRK2; MAPK1; PTPN11; PIK3CA; CREB1;
FOS; PIK3CB; PIK3C3; MAPK8; MAPK3; KRAS; PIK3C2A; RAF1; MAP2K2;
AKT1; PIK3R1; PDPK1; MAP2K1; CDC42; JUN; ATF4.
[0275] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: FXR/RXR
Activation, INS; PPARA; FASN; RXRA; AKT2; SDC1; MAPK8; APOB;
MAPK10; PPARG; MTTP; MAPK9; PPARGC1A; TNF; CREBBP; AKT1; SREBF1;
FGFR4; AKT3; FOXO1.
[0276] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Synaptic Long
Term Potentiation, PRKCE; RAP1A; EP300; PRKCZ; MAPK1; CREB1; PRKC1;
GNAQ; CAMK2A; PRKD1; MAPK3; KRAS; PRKCD; PPP1CC; RAF1; CREBBP;
MAP2K2; MAP2K1; ATF4; PRKCA.
[0277] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Calcium
Signaling, RAP1A; EP300; HDAC4; MAPK1; HDAC5; CREB1; CAMK2A; MYH9;
MAPK3; HDAC2; HDAC7A; HDAC11; HDAC9; HDAC3; CREBBP; CALR; CAMKK2;
ATF4; HDAC6.
[0278] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: EGF
Signaling, ELK1; MAPK1; EGFR; PIK3CA; FOS; PIK3CB; PIK3C3; MAPK8;
MAPK3; PIK3C2A; RAF1; JAK1; PIK3R1; STAT3; MAP2K1; JUN; PRKCA; SRF;
STAT1.
[0279] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Hypoxia
Signaling in the Cardiovascular System, EDN1; PTEN; EP300; NQO1;
UBE21; CREB1; ARNT; HIF1A; SLC2A4; NOS3; TP53; LDHA; AKT1; ATM;
VEGFA; JUN; ATF4; VHL; HSP90AA1.
[0280] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: LPS/IL-1
Mediated Inhibition of RXR Function, IRAK1; MYD88; TRAF6; PPARA;
RXRA; ABCA1, MAPK8; ALDH1A1; GSTP1; MAPK9; ABCB1; TRAF2; TLR4; TNF;
MAP3K7; NR1H2; SREBF1; JUN; IL1R1.
[0281] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: LXR/RXR
Activation, FASN; RXRA; NCOR2; ABCA1; NFKB2; IRF3; RELA; NOS2A;
TLR4; TNF; RELB; LDLR; NR1H2; NFKB1; SREBF1; IL1R1; CCL2; IL6;
MMP9.
[0282] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Amyloid
Processing, PRKCE; CSNK11E; MAPK1; CAPNS1; AKT2; CAPN2; CAPN1;
MAPK3; MAPK13; MAPT; MAPK14; AKT1; PSEN1; CSNK1A1; GSK3B; AKT3;
APP.
[0283] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: IL-4
Signaling, AKT2; PIK3CA; PIK3CB; PIK3C3; IRS1; KRAS; SOCS1; PTPN6;
NR3C1; PIK3C2A; JAK1; AKT1; JAK2; PIK3R1; FRAP1; AKT3; RPS6KB1.
[0284] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Cell Cycle:
G2/M DNA Damage Checkpoint Regulation, EP300; PCAF; BRCA1; GADD45A;
PLK1; BTRC; CHEK1; ATR; CHEK2; YWHAZ; TP53; CDKN1A; PRKDC; ATM;
SFN; CDKN2A.
[0285] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Nitric Oxide
Signaling in the Cardiovascular System, KDR; FLT1; PGF; AKT2;
PIK3CA; PIK3CB; PIK3C3; CAV1; PRKCD; NOS3; PIK3C2A; AKT1; PIK3R1;
VEGFA; AKT3; HSP90AA1.
[0286] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Purine
Metabolism NME2; SMARCA4; MYH9; RRM2; ADAR; EIF2AK4; PKM2; ENTPD1;
RAD51; RRM2B; TJP2; RAD51C; NT5E; POLD1; NME1.
[0287] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: cAMP-mediated
Signaling, RAP1A; MAPK1; GNAS; CREB1; CAMK2A; MAPK3; SRC; RAF1;
MAP2K2; STAT3; MAP2K1; BRAF; ATF4.
[0288] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Mitochondrial
Dysfunction Notch Signaling, SOD2; MAPK8; CASP8; MAPK10; MAPK9;
CASP9; PARK7; PSEN1; PARK2; APP; CASP3 HES1; JAG1; NUMB; NOTCH4;
ADAM17; NOTCH2; PSEN1; NOTCH3; NOTCH1; DLL4.
[0289] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Endoplasmic
Reticulum Stress Pathway, HSPA5; MAPK8; XBP1; TRAF2; ATF6; CASP9;
ATF4; EIF2AK3; CASP3.
[0290] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pyrimidine
Metabolism, NME2; AICDA; RRM2; EIF2AK4; ENTPD1; RRM2B; NT5E; POLD1;
NME1.
[0291] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Parkinson's
Signaling, UCHL1; MAPK8; MAPK13; MAPK14; CASP9; PARK7; PARK2;
CASP3.
[0292] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Cardiac &
Beta Adrenergic Signaling, GNAS; GNAQ; PPP2R1A; GNB2L1; PPP2CA;
PPP1CC; PPP2R5C.
[0293] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Glycolysis/Gluco-neogenesis, HK2; GCK; GPI; ALDH1A1; PKM2; LDHA;
HK1.
[0294] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Interferon
Signaling, IRF1; SOCS1; JAK1; JAK2; IFITM1; STAT1; IFIT3.
[0295] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Sonic
Hedgehog Signaling, ARRB2; SMO; GLI2; DYRK1A; GLI1; GSK3B;
DYRKIB.
[0296] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Glycerophospholipid Metabolism, PLD1; GRN; GPAM; YWHAZ; SPHK1;
SPHK2.
[0297] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Phospholipid
Degradation, PRDX6; PLD1; GRN; YWHAZ; SPHK1; SPHK2.
[0298] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Tryptophan
Metabolism, SIAH2; PRMT5; NEDD4; ALDH1A1; CYP1B1; SIAH1.
[0299] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Lysine
Degradation, SUV39H1; EHMT2; NSD1; SETD7; PPP2R5C.
[0300] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Nucleotide
Excision, ERCC5; ERCC4; XPA; XPC; ERCC1.
[0301] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Repair
Pathway Starch and Sucrose Metabolism, UCHL1; HK2; GCK; GPI;
HK1.
[0302] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Aminosugars
Metabolism, NQO1; HK2; GCK; HK1.
[0303] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Arachidonic
Acid Metabolism, PRDX6; GRN; YWHAZ; CYP1B1.
[0304] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Circadian
Rhythm Signaling, CSNK1E; CREB1; ATF4; NR1D1.
[0305] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Coagulation
System, BDKRB1; F2R; SERPINE1; F3.
[0306] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Dopamine
Receptor Signaling, PPP2R1A; PPP2CA; PPP1CC; PPP2R5C.
[0307] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Glutathione
Metabolism, IDH2; GSTP1; ANPEP; IDH1.
[0308] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Glycerolipid
Metabolism, ALDH1A1; GPAM; SPHK1; SPHK2.
[0309] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Linoleic Acid
Metabolism, PRDX6; GRN; YWHAZ; CYP1B1.
[0310] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Methionine
Metabolism, DNMT1; DNMT3B; AHCY; DNMT3A.
[0311] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pyruvate
Metabolism, GLO1; ALDH1A1; PKM2; LDHA.
[0312] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Arginine and
Proline Metabolism, ALDH1A1; NOS3; NOS2A.
[0313] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Eicosanoid
Signaling, PRDX6; GRN; YWHAZ.
[0314] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Fructose and
Mannose Metabolism, HK2; GCK; HK1.
[0315] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Galactose
Metabolism, HK2; GCK; HK1.
[0316] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Stilbene,
Coumarine and Lignin Biosynthesis, PRDX6; PRDX1; TYR.
[0317] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Antigen
Presentation Pathway, CALR; B2M.
[0318] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Biosynthesis
of Steroids, NQO1; DHCR7.
[0319] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Butanoate
Metabolism, ALDH1A1; NLGN1.
[0320] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Citrate
Cycle, IDH2; IDH1.
[0321] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Fatty Acid
Metabolism, ALDH1A1; CYP1B1.
[0322] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Glycerophospholipid Metabolism, PRDX6; CHKA.
[0323] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Histidine
Metabolism, PRMT5; ALDH1A1.
[0324] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Inositol
Metabolism, ERO1L; APEX1.
[0325] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Metabolism of
Xenobiotics by Cytochrome p450, GSTP1; CYP1B1.
[0326] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Methane
Metabolism, PRDX6; PRDX1.
[0327] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Phenylalanine
Metabolism, PRDX6; PRDX1.
[0328] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Propanoate
Metabolism, ALDH1A1; LDHA.
[0329] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Selenoamino
Acid Metabolism, PRMT5; AHCY.
[0330] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Sphingolipid
Metabolism, SPHK1; SPHK2.
[0331] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include:
Aminophosphonate Metabolism, PRMT5.
[0332] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Androgen and
Estrogen Metabolism, PRMT5.
[0333] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Ascorbate and
Aldarate Metabolism, ALDH1A1.
[0334] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Bile Acid
Biosynthesis, ALDH1A1.
[0335] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Cysteine
Metabolism, LDHA.
[0336] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Fatty Acid
Biosynthesis, FASN.
[0337] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Glutamate
Receptor Signaling, GNB2L1.
[0338] Examples of genes and/or polynucleotides that can be edited
with the guide molecules of this invention include: NRF2-mediated
Oxidative Stress Response, PRDX1.
[0339] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pentose
Phosphate Pathway, GPI.
[0340] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pentose and
Glucuronate Interconversions, UCHL1.
[0341] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Retinol
Metabolism, ALDH1A1.
[0342] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Riboflavin
Metabolism, TYR.
[0343] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Tyrosine
Metabolism, PRMT5, TYR.
[0344] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Ubiquinone
Biosynthesis, PRMT5.
[0345] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Valine,
Leucine and Isoleucine Degradation, ALDH1A1.
[0346] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Glycine,
Serine and Threonine Metabolism, CHKA.
[0347] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Lysine
Degradation, ALDH1A1.
[0348] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pain/Taste,
TRPM5; TRPA1.
[0349] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Pain, TRPM7;
TRPC5; TRPC6; TRPC1; Cnr1; cnr2; Grk2; Trpa1; Pomc; Cgrp; Crf; Pka;
Era; Nr2b; TRPM5; Prkaca; Prkacb; Prkar1a; Prkar2a.
[0350] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Mitochondrial
Function, AIF; CytC; SMAC (Diablo); Aifm-1; Aifm-2.
[0351] Examples of genes for which a mUNA molecule can be used to
express the corresponding peptide or protein include: Developmental
Neurology, BMP-4; Chordin (Chrd); Noggin (Nog); WNT (Wnt2; Wnt2b;
Wnt3a; Wnt4; Wnt5a; Wnt6; Wnt7b; Wnt8b; Wnt9a; Wnt9b; Wnt10a;
Wnt10b; Wnt16); beta-catenin; Dick-1; Frizzled related proteins;
Otx-2; Gbx2; FGF-8; Reelin; Dab1; unc-86 (Pou4f1 or Brn3a); Numb;
Reln.
[0352] mUNA Methods
[0353] In various aspects, this invention provides methods for
synthesis of mUNA messenger UNA oligomer molecules.
[0354] mUNA oligomer molecules of this invention can be synthesized
and isolated using methods disclosed herein, as well as any
pertinent techniques known in the art.
[0355] Some methods for preparing nucleic acids are given in, for
example, Merino, Chemical Synthesis of Nucleoside Analogues,
(2013); Gait, Oligonucleotide synthesis: a practical approach
(1984); Herdewijn, Oligonucleotide Synthesis, Methods in Molecular
Biology, Vol. 288 (2005).
[0356] In some embodiments, a ligase can be used to link a
synthetic oligomer to the 3' end of an RNA molecule or an RNA
transcript to form a mUNA molecule. The synthetic oligomer that is
ligated to the 3' end can provide the functionality of a polyA
tail, and advantageously provide resistance to its removal by
3'-exoribonucleases. The ligated product mUNA molecule can have
increased specific activity and provide increased levels of ectopic
protein expression.
[0357] In certain embodiments, ligated product mUNA molecules of
this invention can be made with an RNA transcript that has native
specificity. The ligated product can be a synthetic molecule that
retains the structure of the RNA transcript at the 5' end to ensure
compatibility with the native specificity.
[0358] In further embodiments, ligated product mUNA molecules of
this invention can be made with an exogenous RNA transcript or
non-natural RNA. The ligated product can be a synthetic molecule
that retains the structure of the RNA.
[0359] In general, the canonical mRNA degradation pathway in cells
includes the steps: (i) the polyA tail is gradually cut back to a
stub by 3' exonucleases, shutting down the looping interaction
required for efficient translation and leaving the cap open to
attack; (ii) decapping complexes remove the 5' cap; (iii) the
unprotected and translationally incompetent residuum of the
transcript is degraded by 5' and 3' exonuclease activity.
[0360] Embodiments of this invention involve new mUNA structures
which can have increased translational activity over a native
transcript. The mUNA molecules can prevent exonucleases from
trimming back the polyA tail in the process of de-adenylation.
[0361] Embodiments of this invention provide structures,
compositions and methods for translatable mUNA molecules.
Embodiments of this invention can provide translatable mUNA
molecules containing one or more UNA monomers and having increased
functional half-life.
[0362] It has been found that ligation of a synthetic oligomer to
the 3' end of an mRNA transcript can surprisingly be accomplished
with high conversion of the mRNA transcript to the ligation
product. The ligase can catalyze the joining of the 3'-hydroxyl
terminus of the RNA transcript to a synthetic oligomer bearing a 5'
monophosphate group. The 3' end of the synthetic oligomer can be
blocked to prevent circularization and concatemerization, while the
presence of a triphosphate or cap moiety at the 5' terminus of the
mRNA transcript can prevent its entry into undesired side
reactions.
[0363] In some embodiments, the yield of conversion of the mRNA
transcript to the ligation product mUNA molecule can be from 70% to
100%. In some embodiments, the yield of conversion of the mRNA
transcript to the ligation product can be 70%, 80%, 90%, 95%, 99%,
or 100%.
[0364] As used herein, the terms polyA tail and polyA oligomer
refer to an oligomer of monomers, wherein the monomers can include
nucleotides based on adenine, UNA monomers, naturally-occurring
nucelotides, modified nucleotides, or nucleotide analogues.
[0365] A modified nucleotide can be base-modified, sugar-modified,
or linkage modified.
[0366] Splint Ligation Methods
[0367] Embodiments of this invention can employ splint ligation to
synthesize mUNA molecules.
[0368] In some aspects, ligation of a tail oligomer to the 3' end
of an RNA molecule can surprisingly be accomplished with high
conversion of the RNA molecule to the ligation product by using a
DNA splint oligomer. Splint ligation of specific RNA molecules can
be done with a DNA ligase and a bridging DNA splint oligomer that
is complementary to the RNAs.
[0369] As used herein, a molecule to which a tail oligomer is added
can be referred to as an acceptor oligomer, and a tail oligomer to
be ligated to an acceptor oligomer can be referred to as a donor
oligomer.
[0370] A donor oligomer of this invention may contain one or more
UNA monomers. In some embodiments, a donor oligomer may be composed
of UNA monomers and adenylate nucleotides.
[0371] A donor oligomer of this invention may include any number of
UNA monomers within its total length.
[0372] An acceptor oligomer of this invention can be a RNA of any
length, an mRNA, or a mammalian gene transcript.
[0373] In some aspects, ligation of a donor oligomer of any length
to the 3' end of an acceptor RNA molecule can surprisingly be
accomplished with high conversion to the ligation product mUNA
molecule by using a DNA splint oligomer.
[0374] In certain embodiments, a DNA splint oligomer can hybridize
to the end of an mRNA having a short polyA tail, anchored in a
specific position based on a region complementary to the end of the
mRNA's 3' UTR. The polyA tail can be about 30 monomers or less in
length. The DNA splint oligomer can incorporate a poly(dT) tail
that overhangs beyond the native polyA tail of the mRNA transcript.
The poly(dT) tail can bring a polyA oligomer into position for
efficient ligation to the synthetic mRNA.
[0375] Embodiments of this invention can employ splint ligation to
introduce UNA monomers, modified nucleotides, or nucleotide
analogues into RNA molecules.
[0376] In certain embodiments, in splint ligation the DNA ligase
can be used to join RNA molecules in an RNA:DNA hybrid.
[0377] In some embodiments, the donor can be from 2 to 120 monomers
in length, or from 3 to 120 monomers, or from 4 to 120 monomers, or
from 5 to 120 monomers, or from 6 to 120 monomers, or longer.
[0378] The splint oligomer can be removed from the ligation product
using a DNAse treatment, which can be required post-IVT to remove
the DNA template for transcription.
[0379] Cohesive End Ligation
[0380] In some embodiments, a wild-type T4 RNA ligase can be used
to join the 3' hydroxyl terminus of an RNA transcript to a tail
oligomer bearing a 5' monophosphate group.
[0381] In further embodiments, a KQ mutant variant of T4 RNA Ligase
2, which requires a pre-adenylated donor, was used to join the 3'
hydroxyl terminus of an RNA transcript to a pre-adenylated tail
oligomer.
[0382] In these embodiments, a preponderance of the tail can
advantageously be incorporated co-transcriptionally in the IVT
synthetic RNA transcript, and the donor oligomer can be
correspondingly shortened.
[0383] Post-Ligation Treatment
[0384] In some aspects, a 3'-exonuclease treatment can be used to
remove the unligated fraction of the product of the ligation
reaction. Examples of a 3'-exonuclease include Exonuclease T,
Ribonuclease R, and analogs thereof.
[0385] In certain embodiments, Ribonuclease R can be used with high
processivity, and the ligation can be insensitive to sequence
content and variations, as well as secondary structure.
[0386] Tail Oligomers
[0387] In some embodiments, the 100% bulk ligation of a tail
oligomer to the 3' end of an RNA has been achieved.
[0388] Donor oligomers of this invention for ligation to the 3' end
of an mRNA may be from 2 to 120 monomers in length, or from 3 to
120 monomers in length, or from 4 to 120 monomers in length, or
from 5 to 120 monomers in length, or longer.
[0389] In further embodiments, a donor oligomer may have a
3'-terminal modification to block circularization or
oligimerization of the synthetic oligomer in ligation reactions.
Examples of a 3'-terminal modification include a 3'-terminal C3
spacer.
[0390] A donor oligomer of this invention may contain one or more
UNA monomers.
[0391] A donor oligomer can include one or more nucleic acid
monomers that are naturally-occurring nucleotides, modified
naturally-occurring nucleotides, or non-naturally-occurring
nucleotides.
[0392] A donor oligomer can include a nucleic acid monomer that is
base-modified, sugar-modified, or linkage modified.
[0393] Pharmaceutical Compositions
[0394] In some aspects, this invention provides pharmaceutical
compositions containing a mUNA oligomeric compound and a
pharmaceutically acceptable carrier.
[0395] A pharmaceutical composition can be capable of local or
systemic administration. In some aspects, a pharmaceutical
composition can be capable of any modality of administration. In
certain aspects, the administration can be intravenous,
subcutaneous, pulmonary, intramuscular, intraperitoneal, dermal,
oral, or nasal administration.
[0396] Embodiments of this invention include pharmaceutical
compositions containing an oligomeric compound in a lipid
formulation.
[0397] In some embodiments, a pharmaceutical composition may
comprise one or more lipids selected from cationic lipids, anionic
lipids, sterols, pegylated lipids, and any combination of the
foregoing.
[0398] In certain embodiments, a pharmaceutical composition can be
substantially free of liposomes.
[0399] In further embodiments, a pharmaceutical composition can
include liposomes or nanoparticles.
[0400] Some examples of lipids and lipid compositions for delivery
of an active molecule of this invention are given in
WO/2015/074085, which is hereby incorporated by reference in its
entirety.
[0401] In additional embodiments, a pharmaceutical composition can
contain an oligomeric compound within a viral or bacterial
vector.
[0402] A pharmaceutical composition of this disclosure may include
carriers, diluents or excipients as are known in the art. Examples
of pharmaceutical compositions and methods are described, for
example, in Remington's Pharmaceutical Sciences, Mack Publishing
Co. (A. R. Gennaro ed. 1985), and Remington, The Science and
Practice of Pharmacy, 21st Edition (2005).
[0403] Examples of excipients for a pharmaceutical composition
include antioxidants, suspending agents, dispersing agents,
preservatives, buffering agents, tonicity agents, and
surfactants.
[0404] An effective dose of an agent or pharmaceutical formulation
of this invention can be an amount that is sufficient to cause
translation of a mUNA molecule in a cell.
[0405] A therapeutically effective dose can be an amount of an
agent or formulation that is sufficient to cause a therapeutic
effect. A therapeutically effective dose can be administered in one
or more separate administrations, and by different routes.
[0406] A therapeutically effective dose, upon administration, can
result in serum levels of an active agent of 1-1000 pg/ml, or
1-1000 ng/ml, or 1-1000 .mu.g/ml, or more.
[0407] A therapeutically effective dose of an active agent in vivo
can be a dose of 0.001-0.01 mg/kg body weight, or 0.01-0.1 mg/kg,
or 0.1-1 mg/kg, or 1-10 mg/kg, or 10-100 mg/kg.
[0408] A therapeutically effective dose of an active agent in vivo
can be a dose of 0.001 mg/kg body weight, or 0.01 mg/kg, or 0.1
mg/kg, or 1 mg/kg, or 2 mg/kg, or 3 mg/kg, or 4 mg/kg, or 5 mg/kg,
or more.
[0409] A subject can be an animal, or a human subject or
patient.
[0410] Base sequences show herein are from left to right, 5' to 3',
unless stated otherwise.
[0411] For the examples below, the mUNA transfection protocol in
vitro was as follows: [0412] 1Plate mouse hepatocyte Hepa1-6 cells
5000 cells per well in 96 well plate at least 8 hours before
transfection. [0413] 2 Replace 90 uL DMEM medium containing 10% FBS
and Non-essential amino acid) adding 90 uL into each well of 96
well plate immediately before beginning the transfection
experiment. [0414] 3 Prepare Messenger Max transfection reagent
(Life Technologies) mUNA complex according to manufacturer's
instruction. [0415] 4 Transfer 10 uL of the complex into a well
containing the cells in the 96-well plate. [0416] 5 Collect the
medium after desired time points and add 100 uL fresh medium into
each well. Medium will be kept at -80.degree. C. until ELISA assay
is performed using the standard manufacturer protocol.
[0417] For the examples below, the mUNA transfection protocol in
vivo was as follows: [0418] 1 The mUNA is formulated with Lipid
nanoparticle (LNP). [0419] 2 Inject the LNP-formulated mUNA (1
mg/kg mUNA) into BL57BL/c mice (4-6 week-old) via standard i.v.
injection in the lateral tail vein. [0420] 3 Collect approximately
50 uL of blood in a Heparin-coated microcentrifuge tube. [0421] 4
Centrifuge at 3,000.times.g for 10 minutes at 4.degree. C. [0422] 5
Transfer the supernatant (plasma) into a fresh microcentrifuge
tube. Plasma will be kept at -80.degree. C. until ELISA assay is
performed using the standard manufacturer protocol.
EXAMPLES
[0423] All of the comparative mUNA and mRNA molecules in the
examples below were synthesized with the 5' cap being a m7GpppGm
cap. Unless otherwise specified, the mUNA molecules in the examples
below contained a 5'-UTR of TEV, and a 3' UTR of xenopus
beta-globin.
Example 1
mUNA Oligomer Producing Human Factor IX in Vivo
[0424] In this example, a translatable mUNA molecule was made and
used for expressing human Factor IX (FT9) in vivo with
advantageously increased efficiency of translation, as compared to
the mRNA of Factor IX. The translatable mUNA molecule expressing
human Factor IX in vivo exhibited activity suitable for use in
methods for ameliorating or treating hemophilia B. In this
embodiment, the translatable mUNA molecule comprised a 5' cap
(m7GpppGm), a 5' UTR of TEV, a F9 CDS, a 3'UTR of xenopus
beta-globin, and a tail region.
[0425] The translation efficiency of this mUNA molecule is shown in
FIG. 1, as compared to the mRNA of F9.
[0426] The mUNA molecule of this embodiment was translated in
C57BL/c mouse to produce human F9.
[0427] FIG. 1 shows that the translation efficiency of this mUNA
molecule was advantageously and surprisingly increased as compared
to the mRNA of F9. In particular, after 55 hours, the translation
efficiency of this mUNA molecule was increased by more than 2-fold
(827/388) as compared to the mRNA of F9.
[0428] Details of the base structure of this translatable mUNA
molecule are as follows:
TABLE-US-00002 (SEQ ID NO: 1)
(m7GpppGm)GGGAAACAUAAGUCAACACAACAUAUACAAAACAAACGAA
UCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUC
UUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUA
GCCAUGGCCCAGCGCGUGAACAUGAUCAUGGCAGAAUCACCAGGCCUCAU
CACCAUCUGCCUUUUAGGAUAUCUACUCAGUGCUGAAUGUACAGUUUUUC
UUGAUCAUGAAAACGCCAACAAAAUUCUGAAUCGGCCAAAGAGGUAUAAU
UCAGGUAAAUUGGAAGAGUUUGUUCAAGGGAACCUUGAGAGAGAAUGUAU
GGAAGAAAAGUGUAGUUUUGAAGAAGCACGAGAAGUUUUUGAAAACACUG
AAAGAACAACUGAAUUUUGGAAGCAGUAUGUUGAUGGAGAUCAGUGUGAG
UCCAAUCCAUGUUUAAAUGGCGGCAGUUGCAAGGAUGACAUUAAUUCCUA
UGAAUGUUGGUGUCCCUUUGGAUUUGAAGGAAAGAACUGUGAAUUAGAUG
UAACAUGUAACAUUAAGAAUGGCAGAUGCGAGCAGUUUUGUAAAAAUAGU
GCUGAUAACAAGGUGGUUUGCUCCUGUACUGAGGGAUAUCGACUUGCAGA
AAACCAGAAGUCCUGUGAACCAGCAGUGCCAUUUCCAUGUGGAAGAGUUU
CUGUUUCACAAACUUCUAAGCUCACCCGUGCUGAGACUGUUUUUCCUGAU
GUGGACUAUGUAAAUUCUACUGAAGCUGAAACCAUUUUGGAUAACAUCAC
UCAAAGCACCCAAUCAUUUAAUGACUUCACUCGGGUUGUUGGUGGAGAAG
AUGCCAAACCAGGUCAAUUCCCUUGGCAGGUUGUUUUGAAUGGUAAAGUU
GAUGCAUUCUGUGGAGGCUCUAUCGUUAAUGAAAAAUGGAUUGUAACUGC
UGCCCACUGUGUUGAAACUGGUGUUAAAAUUACAGUUGUCGCAGGUGAAC
AUAAUAUUGAGGAGACAGAACAUACAGAGCAAAAGCGAAAUGUGAUUCGA
AUUAUUCCUCACCACAACUACAAUGCAGCUAUUAAUAAGUACAACCAUGA
CAUUGCCCUUCUGGAACUGGACGAACCCUUAGUGCUAAACAGCUACGUUA
CACCUAUUUGCAUUGCUGACAAGGAAUACACGAACAUCUUCCUCAAAUUU
GGAUCUGGCUAUGUAAGUGGCUGGGGAAGAGUCUUCCACAAAGGGAGAUC
AGCUUUAGUUCUUCAGUACCUUAGAGUUCCACUUGUUGACCGAGCCACAU
GUCUUCGAUCUACAAAGUUCACCAUCUAUAACAACAUGUUCUGUGCUGGC
UUCCAUGAAGGAGGUAGAGAUUCAUGUCAAGGAGAUAGUGGGGGACCCCA
UGUUACUGAAGUGGAAGGGACCAGUUUCUUAACUGGAAUUAUUAGCUGGG
GUGAAGAGUGUGCAAUGAAAGGCAAAUAUGGAAUAUAUACCAAGGUAUCC
CGGUAUGUCAACUGGAUUAAGGAAAAAACAAAGCUCACUUGACUAGUGAC
UGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGA
GUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCA
AAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAU
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAA AA
Example 2
mUNA Oligomer Producing Human Factor IX in Vitro
[0429] In this example, the translatable mUNA molecule of Example 1
(SEQ ID NO: 1) was made and used for expressing human Factor IX
(F9) in vitro with advantageously increased efficiency of
translation, as compared to the mRNA of Factor IX. The translatable
mUNA molecule expressing human Factor IX exhibited activity
suitable for use in methods for ameliorating or treating hemophilia
B.
[0430] The translation efficiency of this mUNA molecule (SEQ ID NO:
1) is shown in FIG. 2, as compared to the mRNA of F9.
[0431] The mUNA molecule of this embodiment was traslated in mouse
hepatocyte cell line Hepa1-6 to produce human F9.
[0432] FIG. 2 shows that the translation efficiency of this mUNA
molecule was advantageously and surprisingly increased as compared
to the mRNA of F9. In particular, after 48 hours, the translation
efficiency of this mUNA molecule was increased by 5-fold (91/16) as
compared to the mRNA of F9.
Example 3
mUNA Oligomer Producing Human Erythropoietin in Vitro
[0433] In this example, a translatable mUNA molecule was made and
used for expressing human Erythropoietin (EPO) in vitro with
advantageously increased efficiency of translation, as compared to
the mRNA of EPO. The translatable mUNA molecule expressing human
EPO exhibited activity suitable for use in methods for ameliorating
or treating certain anemias, inflammatory bowel disease, and/or
certain myelodysplasias. In this embodiment, the translatable mUNA
molecule comprised a 5' cap (m7GpppGm), a 5' UTR of TEV, a human
EPO CDS, a 3'UTR of xenopus beta-globin, and a tail region.
[0434] The translation efficiency of this mUNA molecule is shown in
FIG. 3, as compared to the mRNA of EPO.
[0435] The mUNA molecule of this embodiment was translated in mouse
hepatocyte cell line Hepa1-6 to produce human EPO.
[0436] FIG. 3 shows that the translation efficiency of this mUNA
molecule was advantageously and surprisingly increased as compared
to the mRNA of F9. In particular, after 48 hours, the translation
efficiency of this mUNA molecule was more than doubled (4500/1784)
as compared to the mRNA of EPO.
[0437] Details of the base structure of this translatable mUNA
molecule are as follows:
TABLE-US-00003 (SEQ ID NO: 2)
(m7GpppGm)GGGAAACAUAAGUCAACACAACAUAUACAAAACAAACGAA
UCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUC
UUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUA
GCCAUGGGGGUGCACGAAUGUCCUGCCUGGCUGUGGCUUCUCCUGUCCCU
GCUGUCGCUCCCUCUGGGCCUCCCAGUCCUGGGCGCCCCACCACGCCUCA
UCUGUGACAGCCGAGUCCUGGAGAGGUACCUCUUGGAGGCCAAGGAGGCC
GAGAAUAUCACGACGGGCUGUGCUGAACACUGCAGCUUGAAUGAGAAUAU
CACUGUCCCAGACACCAAAGUUAAUUUCUAUGCCUGGAAGAGGAUGGAGG
UCGGGCAGCAGGCCGUAGAAGUCUGGCAGGGCCUGGCCCUGCUGUCGGAA
GCUGUCCUGCGGGGCCAGGCCCUGUUGGUCAACUCUUCCCAGCCGUGGGA
GCCCCUGCAGCUGCAUGUGGAUAAAGCCGUCAGUGGCCUUCGCAGCCUCA
CCACUCUGCUUCGGGCUCUGGGAGCCCAGAAGGAAGCCAUCUCCCCUCCA
GAUGCGGCCUCAGCUGCUCCACUCCGAACAAUCACUGCUGACACUUUCCG
CAAACUCUUCCGAGUCUACUCCAAUUUCCUCCGGGGAAAGCUGAAGCUGU
ACACAGGGGAGGCCUGCAGGACAGGGGACAGAUGACUAGUGACUGACUAG
GAUCUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUA
AGCUACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUA
GCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AA
Example 4
mUNA Oligomers Producing Mouse Erythropoietin in Vitro
[0438] In this example, several translatable mUNA molecules were
made and used for expressing mouse Erythropoietin (EPO) in vitro
with advantageously increased efficiency of translation, as
compared to the mRNA of EPO. In this embodiment, the translatable
mUNA molecules each comprised a 5' cap (m7GpppGm), a 5' UTR of TEV,
a mouse EPO CDS, a 3'UTR of xenopus beta-globin, and a tail
region.
[0439] The translation efficiency of these mUNA molecules (#2, 3,
4, 5, 6, 7, 8, 9, 10 and 11) are shown in FIG. 4, as compared to
the mRNA of EPO (#1).
[0440] The mUNA molecules of this embodiment were translated in
mouse hepatocyte cell line Hepa1-6 to produce mouse EPO.
[0441] FIG. 4 shows that the translation efficiency of the mUNA
molecules (#2, 3, 4, 5, 6, 7, 8, 9, 10 and 11) was advantageously
and surprisingly increased as compared to the mRNA of EPO (#1). In
particular, after 72 hours, the translation efficiency of the mUNA
molecules was increased by up to 8-fold (0.203/0.025) as compared
to the mRNA of EPO, and the translation efficiency of every mUNA
molecule (#2, 3, 4, 5, 6, 7, 8, 9, 10 and 11) was increased as
compared to the mRNA of EPO (#1).
[0442] Details of the base structure of the translatable mUNA
molecule #2 are as follows:
TABLE-US-00004 (SEQ ID NO: 3)
(m7GpppGm)GGGAAACAUAAGUCAACACAACAUAUACAAAACAAACGAA
UCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUC
UUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUA
GCCAUGGGGGUGCCCGAACGUCCCACCCUGCUGCUUUUACUCUCCUUGCU
ACUGAUUCCUCUGGGCCUCCCAGUCCUCUGUGCUCCCCCACGCCUCAUCU
GCGACAGUCGAGUUCUGGAGAGGUACAUCUUAGAGGCCAAGGAGGCAGAA
AAUGUCACGAUGGGUUGUGCAGAAGGUCCCAGACUGAGUGAAAAUAUUAC
AGUCCCAGAUACCAAAGUCAACUUCUAUGCUUGGAAAAGAAUGGAGGUGG
AAGAACAGGCCAUAGAAGUUUGGCAAGGCCUGUCCCUGCUCUCAGAAGCC
AUCCUGCAGGCCCAGGCCCUGCUAGCCAAUUCCUCCCAGCCACCAGAGAC
CCUUCAGCUUCAUAUAGACAAAGCCAUCAGUGGUCUACGUAGCCUCACUU
CACUGCUUCGGGUACUGGGAGCUCAGAAGGAAUUGAUGUCGCCUCCAGAU
ACCACCCCACCUGCUCCACUCCGAACACUCACAGUGGAUACUUUCUGCAA
GCUCUUCCGGGUCUACGCCAACUUCCUCCGGGGGAAACUGAAGCUGUACA
CGGGAGAGGUCUGCAGGAGAGGGGACAGGTGACUAGUGACUGACUAGGAU
CUGGUUACCACUAAACCAGCCUCAAGAACACCCGAAUGGAGUCUCUAAGC
UACAUAAUACCAACUUACACUUACAAAAUGUUGUCCCCCAAAAUGUAGCC
AUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUUCUUCACAUAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA{tilde over (
A)}
[0443] Details of the base structure of the translatable mUNA
molecules #3 through #11 that were made are the same as molecule
#2, except that the 3' terminal tail regions, the last 40 monomers
are as follows:
TABLE-US-00005 mUNA molecule #3 (SEQ ID NO: 4)
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAA AA mUNA
molecule #4 (SEQ ID NO: 5) AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAAA AAAA mUNA molecule #5 (SEQ ID NO: 6)
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAA AAAAAA mUNA
molecule #6 (SEQ ID NO: 7) AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAA AAAAAAAA mUNA molecule #7 (SEQ ID NO: 8)
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAA AAAAAAAAAA mUNA
molecule #8 (SEQ ID NO: 9) AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAAAAAA AAAAAAAAAAAA mUNA molecule #9 (SEQ ID NO: 10)
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAA AAAAAAAAAAAAAA mUNA
molecule #10 (SEQ ID NO: 11) AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAA AAAAAAAAAAAAAAAA mUNA molecule #11 (SEQ ID NO: 12)
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAA AAAAAAAAAAAAAAAAAA
Example 5
mUNA Oligomer Producing Human Alpha-1-Antitrypsin in Vivo
[0444] In this example, a translatable mUNA molecule was made and
used for expressing human alpha-1-Antitrypsin in vivo with
advantageously increased efficiency of translation, as compared to
the mRNA of human alpha-1-Antitrypsin. The translatable mUNA
molecule expressing human alpha-1-Antitrypsin exhibited activity
suitable for use in methods for ameliorating or treating
alpha-1-Antitrypsin deficiency. In this embodiment, the
translatable mUNA molecule comprised a 5' cap (m7GpppGm), a 5' UTR
of TEV, a human alpha-1-Antitrypsin CDS, a 3'UTR of xenopus
beta-globin, and a tail region.
[0445] The translation efficiency of this mUNA molecule is shown in
FIG. 5, as compared to the mRNA of human alpha-1-Antitrypsin.
[0446] The mUNA molecule of this embodiment was translated in
C57BL/c mouse to produce human alpha-1-Antitrypsin.
[0447] FIG. 5 shows that the translation efficiency of this mUNA
molecule was advantageously and surprisingly increased as compared
to the mRNA of human alpha-1-Antitrypsin. In particular, after 72
hours, the translation efficiency of this mUNA molecule was
increased by more than 3-fold (87.8/25.4) as compared to the mRNA
of human alpha-1-Antitrypsin.
[0448] Details of the base structure of this translatable mUNA
molecule were as follows:
TABLE-US-00006 (SEQ ID NO: 13)
(m7GpppGm)GGGAAACAUAAGUCAACACAACAUAUACAAAACAAACGAA
UCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUUUC
UUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACGAUA
GCCAUGCCGUCUUCUGUCUCGUGGGGCAUCCUCCUGCUGGCAGGCCUGUG
CUGCCUGGUCCCUGUCUCCCUGGCUGAGGAUCCCCAGGGAGAUGCUGCCC
AGAAGACAGAUACAUCCCACCAUGAUCAGGAUCACCCAACCUUCAACAAG
AUCACCCCCAACCUGGCUGAGUUCGCCUUCAGCCUAUACCGCCAGCUGGC
ACACCAGUCCAACAGCACCAAUAUCUUCUUCUCCCCAGUGAGCAUCGCUA
CAGCCUUUGCAAUGCUCUCCCUGGGGACCAAGGCUGACACUCACGAUGAA
AUCCUGGAGGGCCUGAAUUUCAACCUCACGGAGAUUCCGGAGGCUCAGAU
CCAUGAAGGCUUCCAGGAACUCCUCCGUACCCUCAACCAGCCAGACAGCC
AGCUCCAGCUGACCACCGGCAAUGGCCUGUUCCUCAGCGAGGGCCUGAAG
CUAGUGGAUAAGUUUUUGGAGGAUGUUAAAAAGUUGUACCACUCAGAAGC
CUUCACUGUCAACUUCGGGGACACCGAAGAGGCCAAGAAACAGAUCAACG
AUUACGUGGAGAAGGGUACUCAAGGGAAAAUUGUGGAUUUGGUCAAGGAG
CUUGACAGAGACACAGUUUUUGCUCUGGUGAAUUACAUCUUCUUUAAAGG
CAAAUGGGAGAGACCCUUUGAAGUCAAGGACACCGAGGAAGAGGACUUCC
ACGUGGACCAGGUGACCACCGUGAAGGUGCCUAUGAUGAAGCGUUUAGGC
AUGUUUAACAUCCAGCACUGUAAGAAGCUGUCCAGCUGGGUGCUGCUGAU
GAAAUACCUGGGCAAUGCCACCGCCAUCUUCUUCCUGCCUGAUGAGGGGA
AACUACAGCACCUGGAAAAUGAACUCACCCACGAUAUCAUCACCAAGUUC
CUGGAAAAUGAAGACAGAAGGUCUGCCAGCUUACAUUUACCCAAACUGUC
CAUUACUGGAACCUAUGAUCUGAAGAGCGUCCUGGGUCAACUGGGCAUCA
CUAAGGUCUUCAGCAAUGGGGCUGACCUCUCCGGGGUCACAGAGGAGGCA
CCCCUGAAGCUCUCCAAGGCCGUGCAUAAGGCUGUGCUGACCAUCGACGA
GAAAGGGACUGAAGCUGCUGGGGCCAUGUUUUUAGAGGCCAUACCCAUGU
CUAUCCCCCCCGAGGUCAAGUUCAACAAACCCUUUGUCUUCUUAAUGAUU
GAACAAAAUACCAAGUCUCCCCUCUUCAUGGGAAAAGUGGUGAAUCCCAC
CCAAAAAUAACUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCU
CAAGAACACCCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUU
ACAAAAUGUUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAA
AAGAAAGUUUCUUCACAUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAAAA AAAA
Example 6
mUNA Oligomer Producing Human Erythropoietin in Vivo
[0449] In this example, a translatable mUNA molecule was made and
used for expressing human Erythropoietin (EPO) in vivo with
advantageously increased efficiency of translation, as compared to
the mRNA of EPO. The translatable mUNA molecule expressing human
EPO exhibited activity suitable for use in methods for ameliorating
or treating certain anemias, inflammatory bowel disease, and/or
certain myelodysplasias. In this embodiment, the translatable mUNA
molecule comprised a 5' cap (m7GpppGm), a 5' UTR of TEV, a human
EPO CDS, a 3'UTR of xenopus beta-globin, and a tail region.
[0450] The translation efficiency of this mUNA molecule is shown in
FIG. 6, as compared to the mRNA of EPO.
[0451] The mUNA molecule of this embodiment was translated in
C57BL/c mouse to produce human EPO.
[0452] FIG. 6 shows that the translation efficiency of this mUNA
molecule was advantageously and surprisingly increased as compared
to the mRNA of EPO. In particular, after 72 hours, the translation
efficiency of this mUNA molecule was increased by more than 10-fold
(1517/143) as compared to the mRNA of EPO.
[0453] Details of the base structure of this translatable mUNA
molecule were as follows:
TABLE-US-00007 (SEQ ID NO: 14)
(m7GpppGm)GGGAAACAUAAGUCAACACAACAUAUACAAAACAAACGA
AUCUCAAGCAAUCAAGCAUUCUACUUCUAUUGCAGCAAUUUAAAUCAUU
UCUUUUAAAGCAAAAGCAAUUUUCUGAAAAUUUUCACCAUUUACGAACG
AUAGCCAUGGGGGUGCACGAAUGUCCUGCCUGGCUGUGGCUUCUCCUGU
CCCUGCUGUCGCUCCCUCUGGGCCUCCCAGUCCUGGGCGCCCCACCACG
CCUCAUCUGUGACAGCCGAGUCCUGGAGAGGUACCUCUUGGAGGCCAAG
GAGGCCGAGAAUAUCACGACGGGCUGUGCUGAACACUGCAGCUUGAAUG
AGAAUAUCACUGUCCCAGACACCAAAGUUAAUUUCUAUGCCUGGAAGAG
GAUGGAGGUCGGGCAGCAGGCCGUAGAAGUCUGGCAGGGCCUGGCCCUG
CUGUCGGAAGCUGUCCUGCGGGGCCAGGCCCUGUUGGUCAACUCUUCCC
AGCCGUGGGAGCCCCUGCAGCUGCAUGUGGAUAAAGCCGUCAGUGGCCU
UCGCAGCCUCACCACUCUGCUUCGGGCUCUGGGAGCCCAGAAGGAAGCC
AUCUCCCCUCCAGAUGCGGCCUCAGCUGCUCCACUCCGAACAAUCACUG
CUGACACUUUCCGCAAACUCUUCCGAGUCUACUCCAAUUUCCUCCGGGG
AAAGCUGAAGCUGUACACAGGGGAGGCCUGCAGGACAGGGGACAGAUGA
CUAGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACAC
CCGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUG
UUGUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAG
UUUCUUCACAUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAA
AAAA
Example 7
mUNA Oligomer Producing Human CFTR
[0454] In this example, a translatable mUNA molecule is made for
use in expressing human CFTR in vivo. The translatable mUNA
molecule expressing human CFTR in vivo is suitable for use in
methods for ameliorating or treating cystic fibrosis. In this
embodiment, the translatable mUNA molecule comprises a 5' cap
(m7GpppGm), a 5' UTR of TEV, a CFTR CDS, a 3'UTR of xenopus
beta-globin, and a tail region shown in Example 4.
[0455] Human CFTR is accession NM_000492.3.
Example 8
mUNA Oligomer Producing Human ASL
[0456] In this example, a translatable mUNA molecule is made for
use in expressing human argininosuccinate lyase (ASL) in vivo. The
translatable mUNA molecule expressing human ASL in vivo is suitable
for use in methods for ameliorating or treating ASL deficiency. In
this embodiment, the translatable mUNA molecule comprises a 5' cap
(m7GpppGm), a 5' UTR of TEV, a ASL CDS, a 3'UTR of xenopus
beta-globin, and a tail region shown in Example 4.
[0457] Human ASL is accession NM_001024943.1.
Example 9
mUNA Oligomer Producing Human PAH
[0458] In this example, a translatable mUNA molecule is made for
use in expressing human Phenylalanine-4-hydroxylase (PAH) in vivo.
The translatable mUNA molecule expressing human PAH in vivo is
suitable for use in methods for ameliorating or treating
Phenylketonuria (PKU). In this embodiment, the translatable mUNA
molecule comprises a 5' cap (m7GpppGm), a 5' UTR of TEV, a PAH CDS,
a 3'UTR of xenopus beta-globin, and a tail region shown in Example
4.
[0459] Human PAH is accession NM_000277.1.
Example 10
mUNA Oligomer Producing Human NIS.
[0460] In this example, a translatable mUNA molecule is made for
use in expressing human Sodium/iodide cotransporter (NIS) in vivo.
The translatable mUNA molecule expressing human NIS in vivo is
suitable for use in methods for ameliorating or treating thyroid
disease. In this embodiment, the translatable mUNA molecule
comprises a 5' cap (m7GpppGm), a 5' UTR of TEV, a NIS CDS, a 3'UTR
of xenopus beta-globin, and a tail region shown in Example 4.
[0461] Human NIS is accession BC105047.
Example 11
mUNA Oligomer Producing Human NIS
[0462] In this example, a translatable mUNA molecule is made for
use in expressing human Sodium/iodide cotransporter (NIS) in vivo.
The translatable mUNA molecule expressing human NIS in vivo is
suitable for use in methods for ameliorating or treating thyroid
disease. In this embodiment, the translatable mUNA molecule
comprises a 5' cap (m7GpppGm), a 5' UTR of TEV, a NIS CDS, a 3'UTR
of xenopus beta-globin, and a tail region shown in Example 4.
[0463] Human NIS is accession BC105047.
Example 12
mUNA Oligomer Producing Human Hepcidin
[0464] In this example, a translatable mUNA molecule is made for
use in expressing human Hepcidin in vivo. The translatable mUNA
molecule expressing human Hepcidin in vivo is suitable for use in
methods for ameliorating or treating iron deficiency disease. In
this embodiment, the translatable mUNA molecule comprises a 5' cap
(m7GpppGm), a 5' UTR of TEV, a Hepcidin CDS, a 3'UTR of xenopus
beta-globin, and a tail region shown in Example 4.
[0465] Human Hepcidin is accession NM_021175.3.
Example 13
mUNA Oligomer Expressing Factor IX
[0466] In this example, the structures of mUNA molecules for use in
expressing Factor IX are shown.
[0467] Factor IX (F9) is associated with hemophilia B.
[0468] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human Factor IX. The complete mUNA
molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream of
the sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4 to
12) downstream of the sequence below, each of which corresponds to
the structure of the native mRNA of human Factor IX.
TABLE-US-00008 Human Factor IX is accession NM_000133.3. (SEQ ID
NO: 15) AU{circumflex over (G)}CAGCGCGUGAACAUGAUCAUGGCAGAAUC{tilde
over (A)}CCAGGCCUCAUCACCA UCUGCCUUUUAGG{tilde over
(A)}UAUCUACUCAGUGCUGAAUGUACAGUUUU{tilde over (U)}CUUGA
UCAUGAAAACGCCAACAAAAUUCU{circumflex over
(G)}AAUCGGCCAAAGAGGUAUAAUUCA GGUAA{tilde over
(A)}UUGGAAGAGUUUGUUCAAGGGAACCUUGA{circumflex over (G)}AGAGAAUGUAUGG
AAGAAAAGUGUAGUUU{tilde over (U)}GAAGAAGCACGAGAAGUUUUUGAAAACAC{tilde
over (U)}GA AAGAACAACUGAAUUUUGGAAGCAGUA{tilde over
(U)}GUUGAUGGAGAUCAGUGUGAG UCCAAUCC{tilde over
(A)}UGUUUAAAUGGCGGCAGUUGCAAGGAUGA{circumflex over (C)}AUUAAUUCCU
AUGAAUGUUGGUGUCCCUU{tilde over (U)}GGAUUUGAAGGAAAGAACUGUGAAUUAGA
{tilde over (U)}GUAACAUGUAACAUUAAGAAUGGCAGAUG{circumflex over
(C)}GAGCAGUUUUGUAAAAAU AGUGCUGAUAA{circumflex over
(C)}AAGGUGGUUUGCUCCUGUACUGAGGGAUA{tilde over (U)}CGACUUG
CAGAAAACCAGAAGUCCUGUGA{tilde over (A)}CCAGCAGUGCCAUUUCCAUGUGGAAG
AGU{tilde over (U)}UCUGUUUCACAAACUUCUAAGCUCACCCG{tilde over
(U)}GCUGAGACUGUUUUU CCUGAUGUGGACUA{tilde over
(U)}GUAAAUUCUACUGAAGCUGAAACCAUUUUG{circumflex over (G)}AUA
ACAUCACUCAAAGCACCCAAUCAUU{tilde over (U)}AAUGACUUCACUCGGGUUGUUGG
UGGAGA{tilde over (A)}GAUGCCAAACCAGGUCAAUUCCCUUGGCA{circumflex over
(G)}GUUGUUUUGAAU GGUAAAGUUGAUGCAUU{circumflex over
(C)}UGUGGAGGCUCUAUCGUUAAUGAAAAAUG{circumflex over (G)}A
UUGUAACUGCUGCCCACUGUGUUGAAAC{tilde over (U)}GGUGUUAAAAUUACAGUUGU
CGCAGGUGA{tilde over (A)}CAUAAUAUUGAGGAGACAGAACAUACAGA{circumflex
over (G)}CAAAAGCGA AAUGUGAUUCGAAUUAUUCC{tilde over
(U)}CACCACAACUACAAUGCAGCUAUUAAUA A{circumflex over
(G)}UACAACCAUGACAUUGCCCUUCUGGAACU{circumflex over
(G)}GACGAACCCUUAGUGCU AAACAGCUACGU{tilde over
(U)}ACACCUAUUUGCAUUGCUGACAAGGAAUA{circumflex over (C)}ACGAAC
AUCUUCCUCAAAUUUGGAUCUGG{circumflex over
(C)}UAUGUAAGUGGCUGGGGAAGAGUCU UCCA{circumflex over
(C)}AAAGGGAGAUCAGCUUUAGUUCUUCAGUA{circumflex over
(C)}CUUAGAGUUCCACU UGUUGACCGAGCCAC{tilde over
(A)}UGUCUUCGAUCUACAAAGUUCACCAUCUA{tilde over (U)}AAC
AACAUGUUCUGUGCUGGCUUCCAUGA{tilde over (A)}GGAGGUAGAGAUUCAUGUCAAG
GAGAUAG{tilde over (U)}GGGGGACCCCAUGUUACUGAAGUGGAAGG{circumflex
over (G)}ACCAGUUUCUU AACUGGAAUUAUUAGCUG{circumflex over
(G)}GGUGAAGAGUGUGCAAUGAAAGGCAAAUA{tilde over (U)}
GGAAUAUAUACCAAGGUAUCCCGGUAUGU{circumflex over
(C)}AACUGGAUUAAGGAAAAAA CAAAGCUCAC{tilde over (U)}UAA (SEQ ID NO:
16) A{tilde over (U)}{circumflex over (G)}{circumflex over
(C)}AGCGCGUGAACAUGAUCAUGGCAGAAUCACCAGGCCUCAUCACCA
UCUGCCUUUUAGGAUAUCUACUCAGUGCUGAAUGUACAGUUUUUCUUGA
UCAUGAAAACGCCAACAAAAUUCUGAAUCGGCCAAAGAGGUAUAAUUCA
GGUAAAUUGGAAGAGUUUGUUCAAGGGAACCUUGAGAGAGAAUGUAUGG
AAGAAAAGUGUAGUUUUGAAGAAGCACGAGAAGUUUUUGAAAACACUGA
AAGAACAACUGAAUUUUGGAAGCAGUAUGUUGAUGGAGAUCAGUGUGAG
UCCAAUCCAUGUUUAAAUGGCGGCAGUUGCAAGGAUGACAUUAAUUCCU
AUGAAUGUUGGUGUCCCUUUGGAUUUGAAGGAAAGAACUGUGAAUUAGA
UGUAACAUGUAACAUUAAGAAUGGCAGAUGCGAGCAGUUUUGUAAAAAU
AGUGCUGAUAACAAGGUGGUUUGCUCCUGUACUGAGGGAUAUCGACUUG
CAGAAAACCAGAAGUCCUGUGAACCAGCAGUGCCAUUUCCAUGUGGAAG
AGUUUCUGUUUCACAAACUUCUAAGCUCACCCGUGCUGAGACUGUUUUU
CCUGAUGUGGACUAUGUAAAUUCUACUGAAGCUGAAACCAUUUUGGAUA
CACAUCACUCAAAGCACCCAAUCAUUUAAUGACUUCACUCGGGUUGUUG
GUGGAGAAGAUGCCAAACCAGGUCAAUUCCCUUGGCAGGUUGUUUUGAA
UGGUAAAGUUGAUGCAUUCUGUGGAGGCUCUAUGUUAAUGAAAAAUGGA
UUGUAACUGCUGCCCACUGUGUUGAAACUGGUGUUAAAAUUACAGUUGU
CGCAGGUGAACAUAAUAUUGAGGAGACAGAACAUACAGAGCAAAAGCGA
AAUGUGAUUCGAAUUAUUCCUCACCACAACUACAAUGCAGCUAUUAAUA
AGUACAACCAUGACAUUGCCCUUCUGGAACUGGACGAACCCUUAGUGCU
AAACAGCUACGUUACACCUAUUUGCAUUGCUGACAAGGAAUACACGAAC
AUCUUCCUCAAAUUUGGAUCUGGCUAUGUAAGUGGCUGGGGAAGAGUCU
UCCACAAAGGGAGAUCAGCUUUAGUUCUUCAGUACCUUAGAGUUCCACU
UGUUGACCGAGCCACAUGUCUUCGAUCUACAAAGUUCACCAUCUAUAAC
AACAUGUUCUGUGCUGGCUUCCAUGAAGGAGGUAGAGAUUCAUGUCAAG
GAGAUAGUGGGGGACCCCAUGUUACUGAAGUGGAAGGGACCAGUUUCUU
AACUGGAAUUAUUAGCUGGGGUGAAGAGUGUGCAAUGAAAGGCAAAUAU
GGAAUAUAUACCAAGGUAUCCCGGUAUGUCAACUGGAUUAAGGAAAAAA CAAAGCUCAC{tilde
over (U)}{tilde over (U)}{tilde over (A)}A (SEQ ID NO: 17) A{tilde
over (U)}GCAGCGCG{tilde over (U)}GAACA{tilde over (U)}GA{tilde over
(U)}CAGGCAGAA{tilde over (U)}CACCAGGCC{tilde over (U)}CA{tilde over
(U)}CACCA{tilde over (U)} C{tilde over (U)}GCC{tilde over
(U)}{tilde over (U)}{tilde over (U)}{tilde over (U)}AGGA{tilde over
(U)}A{tilde over (U)}C{tilde over (U)}AC{tilde over (U)}CAG{tilde
over (U)}GC{tilde over (U)}GAA{tilde over (U)}G{tilde over
(U)}ACAG{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde over
(U)}{tilde over (U)}C{tilde over (U)}{tilde over (U)}GA{tilde over
(U)} CA{tilde over (U)}GAAAACGCCAACAAAA{tilde over (U)}{tilde over
(U)}C{tilde over (U)}GAA{tilde over (U)}CGGCCAAAGAGG{tilde over
(U)}A{tilde over (U)}AA{tilde over (U)}{tilde over (U)}CAG G{tilde
over (U)}AAA{tilde over (U)}{tilde over (U)}GGAAGAG{tilde over
(U)}{tilde over (U)}{tilde over (U)}G{tilde over (U)}{tilde over
(U)}CAAGGGAACC{tilde over (U)}{tilde over (U)}GAGAGAGAA{tilde over
(U)}G{tilde over (U)}A{tilde over (U)}GGA AGAAAAG{tilde over
(U)}G{tilde over (U)}AG{tilde over (U)}{tilde over (U)}{tilde over
(U)}{tilde over (U)}GAAGAAGCACGAGAAG{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}{tilde over (U)}GAAAACAC{tilde
over (U)}GAA AGAACAAC{tilde over (U)}GAA{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}GGAAGCAG{tilde over (U)}A{tilde
over (U)}G{tilde over (U)}{tilde over (U)}GA{tilde over
(U)}GGAGA{tilde over (U)}CAG{tilde over (U)}G{tilde over
(U)}GAG{tilde over (U)} CCAA{tilde over (U)}CCA{tilde over
(U)}G{tilde over (U)}{tilde over (U)}{tilde over (U)}AAA{tilde over
(U)}GGCGGCAG{tilde over (U)}{tilde over (U)}GCAAGGA{tilde over
(U)}GACA{tilde over (U)}{tilde over (U)}AA{tilde over (U)}{tilde
over (U)}CC{tilde over (U)}A {tilde over (U)}GAA{tilde over
(U)}G{tilde over (U)}{tilde over (U)}GG{tilde over (U)}G{tilde over
(U)}CCC{tilde over (U)}{tilde over (U)}{tilde over (U)}GGA{tilde
over (U)}{tilde over (U)}{tilde over (U)}GAAGGAAAGAAC{tilde over
(U)}G{tilde over (U)}GAA{tilde over (U)}{tilde over (U)}AGA{tilde
over (U)} G{tilde over (U)}AACA{tilde over (U)}G{tilde over
(U)}AACA{tilde over (U)}{tilde over (U)}AAGAA{tilde over
(U)}GGCAGA{tilde over (U)}GCGAGCAG{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}G{tilde over (U)}AAAAA{tilde
over (U)}A G{tilde over (U)}GC{tilde over (U)}GA{tilde over
(U)}AACAAGG{tilde over (U)}GG{tilde over (U)}{tilde over (U)}{tilde
over (U)}GC{tilde over (U)}CC{tilde over (U)}G{tilde over
(U)}AC{tilde over (U)}GAGGGA{tilde over (U)}A{tilde over
(U)}CGAC{tilde over (U)}{tilde over (U)}GC AGAAAACCAGAAG{tilde over
(U)}CC{tilde over (U)}G{tilde over (U)}GAACCAGCAG{tilde over
(U)}GCCA{tilde over (U)}{tilde over (U)}{tilde over (U)}CCA{tilde
over (U)}G{tilde over (U)}GGAAGA G{tilde over (U)}{tilde over
(U)}{tilde over (U)}C{tilde over (U)}G{tilde over (U)}{tilde over
(U)}{tilde over (U)}CACAAAC{tilde over (U)}{tilde over (U)}C{tilde
over (U)}AAGC{tilde over (U)}CACCCG{tilde over (U)}GC{tilde over
(U)}GAGAC{tilde over (U)}G{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}{tilde over (U)}C C{tilde over
(U)}GA{tilde over (U)}G{tilde over (U)}GGAC{tilde over (U)}A{tilde
over (U)}G{tilde over (U)}AAA{tilde over (U)}{tilde over
(U)}C{tilde over (U)}AC{tilde over (U)}GAAGC{tilde over
(U)}GAAACCA{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}GGA{tilde over (U)}AA CA{tilde over (U)}CAC{tilde over
(U)}CAAAGCACCCAA{tilde over (U)}CA{tilde over (U)}{tilde over
(U)}{tilde over (U)}AA{tilde over (U)}GAC{tilde over (U)}{tilde
over (U)}CAC{tilde over (U)}CGGG{tilde over (U)}{tilde over
(U)}G{tilde over (U)}{tilde over (U)}GG{tilde over (U)}
GGAGAAGA{tilde over (U)}GCCAAACCAGG{tilde over (U)}CAA{tilde over
(U)}{tilde over (U)}CCC{tilde over (U)}{tilde over (U)}GGCAGG{tilde
over (U)}{tilde over (U)}G{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}GAA{tilde over (U)}G G{tilde over
(U)}AAAG{tilde over (U)}{tilde over (U)}GA{tilde over (U)}GCA{tilde
over (U)}{tilde over (U)}C{tilde over (U)}G{tilde over
(U)}GGAGGC{tilde over (U)}C{tilde over (U)}A{tilde over
(U)}CG{tilde over (U)}{tilde over (U)}AA{tilde over
(U)}GAAAAA{tilde over (U)}GGA{tilde over (U)} {tilde over
(U)}G{tilde over (U)}AAC{tilde over (U)}GC{tilde over
(U)}GCCCAC{tilde over (U)}G{tilde over (U)}G{tilde over (U)}{tilde
over (U)}GAAAC{tilde over (U)}GG{tilde over (U)}G{tilde over
(U)}{tilde over (U)}AAAA{tilde over (U)}{tilde over (U)}ACAG{tilde
over (U)}{tilde over (U)}G{tilde over (U)}C GCAGG{tilde over
(U)}GAACA{tilde over (U)}AA{tilde over (U)}A{tilde over (U)}{tilde
over (U)}GAGGAGACAGAACA{tilde over (U)}ACAGAGCAAAAGCGAA A{tilde
over (U)}G{tilde over (U)}GA{tilde over (U)}{tilde over
(U)}CGAA{tilde over (U)}{tilde over (U)}A{tilde over (U)}{tilde
over (U)}CC{tilde over (U)}CACCACAAC{tilde over (U)}ACAA{tilde over
(U)}GCAGC{tilde over (U)}A{tilde over (U)}{tilde over (U)}AA{tilde
over (U)}AA G{tilde over (U)}ACAACCA{tilde over (U)}GACA{tilde over
(U)}{tilde over (U)}GCCC{tilde over (U)}{tilde over (U)}C{tilde
over (U)}GGAAC{tilde over (U)}GGACGAACCC{tilde over (U)}{tilde over
(U)}AG{tilde over (U)}GC{tilde over (U)}A AACAGC{tilde over
(U)}ACG{tilde over (U)}{tilde over (U)}ACACC{tilde over (U)}A{tilde
over (U)}{tilde over (U)}{tilde over (U)}GCA{tilde over (U)}{tilde
over (U)}GC{tilde over (U)}GACAAGGAA{tilde over (U)}ACACGAACA
{tilde over (U)}C{tilde over (U)}{tilde over (U)}CC{tilde over
(U)}CAAA{tilde over (U)}{tilde over (U)}{tilde over (U)}GGA{tilde
over (U)}C{tilde over (U)}GGC{tilde over (U)}A{tilde over
(U)}G{tilde over (U)}AAG{tilde over (U)}GGC{tilde over
(U)}GGGGAAGAG{tilde over (U)}C{tilde over (U)}{tilde over (U)}
CCACAAAGGGAGA{tilde over (U)}CAGC{tilde over (U)}{tilde over
(U)}{tilde over (U)}AG{tilde over (U)}{tilde over (U)}C{tilde over
(U)}{tilde over (U)}CAG{tilde over (U)}ACC{tilde over (U)}{tilde
over (U)}AGAG{tilde over (U)}{tilde over (U)}CCAC{tilde over
(U)}{tilde over (U)} G{tilde over (U)}{tilde over
(U)}GACCGAGCCACA{tilde over (U)}G{tilde over (U)}C{tilde over
(U)}{tilde over (U)}CGA{tilde over (U)}C{tilde over
(U)}ACAAAG{tilde over (U)}{tilde over (U)}CACCA{tilde over
(U)}C{tilde over (U)}A{tilde over (U)}AACA ACA{tilde over
(U)}G{tilde over (U)}{tilde over (U)}C{tilde over (U)}G{tilde over
(U)}GC{tilde over (U)}GGC{tilde over (U)}{tilde over (U)}CCA{tilde
over (U)}GAAGGAGG{tilde over (U)}AGAGA{tilde over (U)}{tilde over
(U)}CA{tilde over (U)}G{tilde over (U)}CAAGG AGA{tilde over
(U)}AG{tilde over (U)}GGGGGACCCCA{tilde over (U)}G{tilde over
(U)}{tilde over (U)}AC{tilde over (U)}GAAG{tilde over
(U)}GGAAGGGACCAG{tilde over (U)}{tilde over (U)}{tilde over
(U)}C{tilde over (U)}{tilde over (U)}A AC{tilde over (U)}GGAA{tilde
over (U)}{tilde over (U)}A{tilde over (U)}{tilde over (U)}AGC{tilde
over (U)}GGGG{tilde over (U)}GAAGAG{tilde over (U)}G{tilde over
(U)}GCAA{tilde over (U)}GAAAGGCAAA{tilde over (U)}A{tilde over
(U)}G GAA{tilde over (U)}A{tilde over (U)}A{tilde over
(U)}ACCAAGG{tilde over (U)}A{tilde over (U)}CCCGG{tilde over
(U)}A{tilde over (U)}G{tilde over (U)}CAAC{tilde over (U)}GGA{tilde
over (U)}{tilde over (U)}AAGGAAAAAAC AAAGC{tilde over (U)}CAC{tilde
over (U)}{tilde over (U)}AA
Example 14
mUNA Oligomer Expressing Alpha-1-Antitrypsin
[0469] In this example, the structures of mUNA molecules for use in
expressing alpha-1-Antitrypsin are shown.
[0470] Alpha-1-Antitrypsin is associated with alpha-1-Antitrypsin
deficiency disease, cystic fibrosis, interstitial lung disease, and
pulmonary arterial hypertension.
[0471] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of alpha-1-Antitrypsin. The complete mUNA
molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream of
the sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4 to
12) downstream of the sequence below, each of which corresponds to
the structure of the native mRNA of alpha-1-Antitrypsin.
TABLE-US-00009 Human alpha-1-antitrypsin mRNA is accession
NM_000295.4. (SEQ ID NO: 18) AU{circumflex over
(G)}CCGUCUUCUGUCUCGUGGGGCAUCCUCCU{circumflex over
(G)}CUGGCAGGCCUGUGCU GCCUGGUCCCUGU{circumflex over
(C)}UCCCUGGCUGAGGAUCCCCAGGGAGAUGC{tilde over (U)}GCCCA
GAAGACAGAUACAUCCCACCAUGA{tilde over (U)}CAGGAUCACCCAACCUUCAACAAG
AUCA{circumflex over (C)}CCCCAACCUGGCUGAGUUCGCCUUCAGCCU{tilde over
(A)}UACCGCCAGCUGG CACACCAGUCCAACAG{circumflex over
(C)}ACCAAUAUCUUCUUCUCCCCAGUGAGCAU{circumflex over (C)}GC
UACAGCCUUUGCAAUGCUCUCCCUGGG{circumflex over
(G)}ACCAAGGCUGACACUCACGAU GAAAUCCU{circumflex over
(G)}GAGGGCCUGAAUUUCAACCUCACGGAGAU{tilde over (U)}CCGGAGGCUC
AGAUCCAUGAAGGCUUCCA{circumflex over
(G)}GAACUCCUCCGUACCCUCAACCAGCCAGA {circumflex over
(C)}AGCCAGCUCCAGCUGACCACCGGCAAUGG{circumflex over
(C)}CUGUUCCUCAGCGAGGGC CUGAAGCUAGU{circumflex over
(G)}GAUAAGUUUUUGGAGGAUGUUAAAAAGUU{circumflex over (G)}UACCACU
CAGAAGCCUUCACUGUCAACUU{circumflex over
(C)}GGGGACACCGAAGAGGCCAAGAAACA GAU{circumflex over
(C)}AACGAUUACGUGGAGAAGGGUACUCAAGG{circumflex over
(G)}AAAAUUGUGGAUUUG GUCAAGGAGCUUGA{circumflex over
(C)}AGAGACACAGUUUUUGCUCUGGUGAAUUA{circumflex over (C)}AUCU
UCUUUAAAGGCAAAUGGGAGAGACC{circumflex over
(C)}UUUGAAGUCAAGGACACCGAGGA AGAGGA{circumflex over
(C)}UUCCACGUGGACCAGGUGACCACCGUGAA{circumflex over (G)}GUGCCUAUGAUG
AAGCGUUUAGGCAUGUU{tilde over
(U)}AACAUCCAGCACUGUAAGAAGCUGUCCAG{circumflex over (C)}U
GGGUGCUGCUGAUGAAAUACCUGGGCAA{tilde over (U)}GCCACCGCCAUCUUCUUCCU
GCCUGAUGA{circumflex over
(G)}GGGAAACUACAGCACCUGGAAAAUGAACU{circumflex over (C)}ACCCACGAU
AUCAUCACCAAGUUCCUGGA{tilde over (A)}AAUGAAGACAGAAGGUCUGCCAGCUUAC
A{tilde over (U)}UUACCCAAACUGUCCAUUACUGGAACCUA{tilde over
(U)}GAUCUGAAGAGCGUCCU GGGUCAACUGGG{circumflex over
(C)}AUCACUAAGGUCUUCAGCAAUGGGGCUGA{circumflex over (C)}CUCUCC
GGGGUCACAGAGGAGGCACCCCU{circumflex over
(G)}AAGCUCUCCAAGGCCGUGCAUAAGG CUGU{circumflex over
(G)}CUGACCAUCGACGAGAAAGGGACUGAAGC{tilde over (U)}GCUGGGGCCAUGUU
UUUAGAGGCCAUACC{circumflex over
(C)}AUGUCUAUCCCCCCCGAGGUCAAGUUCAA{circumflex over (C)}AAA
CCCUUUGUCUUCUUAAUGAUUGAACA{tilde over (A)}AAUACCAAGUCUCCCCUCUUCA
UGGGAAA{tilde over (A)}GUGGUGAAUCCCACCCAAAAAU{tilde over (A)}A (SEQ
ID NO: 19) A{tilde over (U)}{circumflex over (G)}{circumflex over
(C)}CGUCUUCUGUCUCGUGGGGCAUCCUCCUGCUGGCAGGCCUGUGCU
GCCUGGUCCCUGUCUCCCUGGCUGAGGAUCCCCAGGGAGAUGCUGCCCA
GAAGACAGAUACAUCCCACCAUGAUCAGGAUCACCCAACCUUCAACAAG
AUCACCCCCAACCUGGCUGAGUUCGCCUUCAGCCUAUACCGCCAGCUGG
CACACCAGUCCAACAGCACCAAUAUCUUCUUCUCCCCAGUGAGCAUCGC
UACAGCCUUUGCAAUGCUCUCCCUGGGGACCAAGGCUGACACUCACGAU
GAAAUCCUGGAGGGCCUGAAUUUCAACCUCACGGAGAUUCCGGAGGCUC
AGAUCCAUGAAGGCUUCCAGGAACUCCUCCGUACCCUCAACCAGCCAGA
CAGCCAGCUCCAGCUGACCACCGGCAAUGGCCUGUUCCUCAGCGAGGGC
CUGAAGCUAGUGGAUAAGUUUUUGGAGGAUGUUAAAAAGUUGUACCACU
CAGAAGCCUUCACUGUCAACUUCGGGGACACCGAAGAGGCCAAGAAACA
GAUCAACGAUUACGUGGAGAAGGGUACUCAAGGGAAAAUUGUGGAUUUG
GUCAAGGAGCUUGACAGAGACACAGUUUUUGCUCUGGUGAAUUACAUCU
UCUUUAAAGGCAAAUGGGAGAGACCCUUUGAAGUCAAGGACACCGAGGA
AGAGGACUUCCACGUGGACCAGGUGACCACCGUGAAGGUGCCUAUGAUG
AAGCGUUUAGGCAUGUUUAACAUCCAGCACUGUAAGAAGCUGUCCAGCU
GGGUGCUGCUGAUGAAAUACCUGGGCAAUGCCACCGCCAUCUUCUUCCU
GCCUGAUGAGGGGAAACUACAGCACCUGGAAAAUGAACUCACCCACGAU
AUCAUCACCAAGUUCCUGGAAAAUGAAGACAGAAGGUCUGCCAGCUUAC
AUUUACCCAAACUGUCCAUUACUGGAACCUAUGAUCUGAAGAGCGUCCU
GGGUCAACUGGGCAUCACUAAGGUCUUCAGCAAUGGGGCUGACCUCUCC
GGGGUCACAGAGGAGGCACCCCUGAAGCUCUCCAAGGCCGUGCAUAAGG
CUGUGCUGACCAUCGACGAGAAAGGGACUGAAGCUGCUGGGGCCAUGUU
UUUAGAGGCCAUACCCAUGUCUAUCCCCCCCGAGGUCAAGUUCAACAAA
CCCUUUGUCUUCUUAAUGAUUGAACAAAAUACCAAGUCUCCCCUCUUCA
UGGGAAAAGUGGUGAAUCCCACCCAAAA{tilde over (A)}{tilde over (U)}{tilde
over (A)}A (SEQ ID NO: 20) A{tilde over (U)}GCCG{tilde over
(U)}C{tilde over (U)}{tilde over (U)}C{tilde over (U)}G{tilde over
(U)}C{tilde over (U)}CG{tilde over (U)}GGGGCA{tilde over
(U)}CC{tilde over (U)}CC{tilde over (U)}GC{tilde over
(U)}GGCAGGCC{tilde over (U)}G{tilde over (U)}GC{tilde over (U)}
GCC{tilde over (U)}GG{tilde over (U)}CCC{tilde over (U)}G{tilde
over (U)}C{tilde over (U)}CCC{tilde over (U)}GGC{tilde over
(U)}GAGGA{tilde over (U)}CCCCAGGGAGA{tilde over (U)}GC{tilde over
(U)}GCCCA GAAGACAGA{tilde over (U)}ACA{tilde over (U)}CCCACCA{tilde
over (U)}GA{tilde over (U)}CAGGA{tilde over (U)}CACCCAACC{tilde
over (U)}{tilde over (U)}CAACAAG A{tilde over (U)}CACCCCCAACC{tilde
over (U)}GGC{tilde over (U)}GAG{tilde over (U)}{tilde over
(U)}CGCC{tilde over (U)}{tilde over (U)}CAGCC{tilde over
(U)}A{tilde over (U)}ACCGCCAGC{tilde over (U)}GG CACACCAG{tilde
over (U)}CCAACAGCACCAA{tilde over (U)}A{tilde over (U)}C{tilde over
(U)}{tilde over (U)}C{tilde over (U)}{tilde over (U)}C{tilde over
(U)}CCCCAG{tilde over (U)}GAGCA{tilde over (U)}CGC {tilde over
(U)}ACAGCC{tilde over (U)}{tilde over (U)}{tilde over
(U)}GCAA{tilde over (U)}GC{tilde over (U)}C{tilde over
(U)}CCC{tilde over (U)}GGGGACCAAGGC{tilde over (U)}GACAC{tilde over
(U)}CACGA{tilde over (U)} GAAA{tilde over (U)}CC{tilde over
(U)}GGAGGGCC{tilde over (U)}GAA{tilde over (U)}{tilde over
(U)}{tilde over (U)}CAACC{tilde over (U)}CACGGAGA{tilde over
(U)}{tilde over (U)}CCGGAGGC{tilde over (U)}C AGA{tilde over
(U)}CCA{tilde over (U)}GAAGGC{tilde over (U)}{tilde over
(U)}CCAGGAAC{tilde over (U)}CC{tilde over (U)}CCG{tilde over
(U)}ACCC{tilde over (U)}CAACCAGCCAGA CAGCCAGC{tilde over
(U)}CCAGC{tilde over (U)}GACCACCGGCAA{tilde over (U)}GGCC{tilde
over (U)}G{tilde over (U)}{tilde over (U)}CC{tilde over
(U)}CAGCGAGGGC C{tilde over (U)}GAAGC{tilde over (U)}AG{tilde over
(U)}GGA{tilde over (U)}AAG{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}{tilde over (U)}GGAGGA{tilde over
(U)}G{tilde over (U)}{tilde over (U)}AAAAAG{tilde over (U)}{tilde
over (U)}G{tilde over (U)}ACCAC{tilde over (U)} CAGAAGCC{tilde over
(U)}{tilde over (U)}CAC{tilde over (U)}G{tilde over (U)}CAAC{tilde
over (U)}{tilde over (U)}CGGGGACACCGAAGAGGCCAAGAAACA GA{tilde over
(U)}CAACGA{tilde over (U)}{tilde over (U)}ACG{tilde over
(U)}GGAGAAGGG{tilde over (U)}AC{tilde over (U)}CAAGGGAAAA{tilde
over (U)}{tilde over (U)}G{tilde over (U)}GGA{tilde over (U)}{tilde
over (U)}{tilde over (U)}G G{tilde over (U)}CAAGGAGC{tilde over
(U)}{tilde over (U)}GACAGAGACACAG{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}{tilde over (U)}GC{tilde over
(U)}C{tilde over (U)}GG{tilde over (U)}GAA{tilde over (U)}{tilde
over (U)}ACA{tilde over (U)}C{tilde over (U)} {tilde over
(U)}C{tilde over (U)}{tilde over (U)}{tilde over
(U)}AAAGGCAAA{tilde over (U)}GGGAGAGACCC{tilde over (U)}{tilde over
(U)}{tilde over (U)}GAAG{tilde over (U)}CAAGGACACCGAGGA
AGAGGAC{tilde over (U)}{tilde over (U)}CCACG{tilde over
(U)}GGACCAGG{tilde over (U)}GACCACCG{tilde over (U)}GAAGG{tilde
over (U)}GCC{tilde over (U)}A{tilde over (U)}GA{tilde over (U)}G
AAGCG{tilde over (U)}{tilde over (U)}{tilde over (U)}AGGCA{tilde
over (U)}G{tilde over (U)}{tilde over (U)}{tilde over
(U)}AACA{tilde over (U)}CCAGCAC{tilde over (U)}G{tilde over
(U)}AAGAAGC{tilde over (U)}G{tilde over (U)}CCAGC{tilde over (U)}
GGG{tilde over (U)}GC{tilde over (U)}GC{tilde over (U)}GA{tilde
over (U)}GAAA{tilde over (U)}ACC{tilde over (U)}GGGCAA{tilde over
(U)}GCCACCGCCA{tilde over (U)}C{tilde over (U)}{tilde over
(U)}C{tilde over (U)}{tilde over (U)}CC{tilde over (U)} GCC{tilde
over (U)}GA{tilde over (U)}GAGGGGAAAC{tilde over (U)}ACAGCACC{tilde
over (U)}GGAAAA{tilde over (U)}GAAC{tilde over (U)}CACCCACGA{tilde
over (U)} A{tilde over (U)}CA{tilde over (U)}CACCAAG{tilde over
(U)}{tilde over (U)}CC{tilde over (U)}GGAAAA{tilde over
(U)}GAAGACAGAAGG{tilde over (U)}C{tilde over (U)}GCCAGC{tilde over
(U)}{tilde over (U)}AC A{tilde over (U)}{tilde over (U)}{tilde over
(U)}ACCCAAAC{tilde over (U)}G{tilde over (U)}CCA{tilde over
(U)}{tilde over (U)}AC{tilde over (U)}GGAACC{tilde over (U)}A{tilde
over (U)}GA{tilde over (U)}C{tilde over (U)}GAAGAGCG{tilde over
(U)}CC{tilde over (U)} GGG{tilde over (U)}CAAC{tilde over
(U)}GGGCA{tilde over (U)}CAC{tilde over (U)}AAGG{tilde over
(U)}C{tilde over (U)}{tilde over (U)}CAGCAA{tilde over
(U)}GGGGC{tilde over (U)}GACC{tilde over (U)}C{tilde over (U)}CC
GGGG{tilde over (U)}CACAGAGGAGGCACCCC{tilde over (U)}GAAGC{tilde
over (U)}C{tilde over (U)}CCAAGGCCG{tilde over (U)}GCA{tilde over
(U)}AAGG C{tilde over (U)}G{tilde over (U)}GC{tilde over
(U)}GACCA{tilde over (U)}CGACGAGAAAGGGAC{tilde over (U)}GAAGC{tilde
over (U)}GC{tilde over (U)}GGGGCCA{tilde over (U)}G{tilde over
(U)}{tilde over (U)} {tilde over (U)}{tilde over (U)}{tilde over
(U)}AGAGGCCA{tilde over (U)}ACCCA{tilde over (U)}G{tilde over
(U)}C{tilde over (U)}A{tilde over (U)}CCCCCCCGAGG{tilde over
(U)}CAAG{tilde over (U)}{tilde over (U)}CAACAAA CCC{tilde over
(U)}{tilde over (U)}{tilde over (U)}G{tilde over (U)}C{tilde over
(U)}{tilde over (U)}C{tilde over (U)}{tilde over (U)}AA{tilde over
(U)}GA{tilde over (U)}{tilde over (U)}GAACAAAA{tilde over
(U)}ACCAAG{tilde over (U)}C{tilde over (U)}CCCC{tilde over
(U)}C{tilde over (U)}{tilde over (U)}CA {tilde over
(U)}GGGAAAAG{tilde over (U)}GG{tilde over (U)}GAA{tilde over
(U)}CCCACCCAAAAA{tilde over (U)}AA
Example 15
mUNA Oligomer Expressing Alpha-1-Antitrypsin
[0472] In this example, the structures of mUNA molecules for use in
expressing alpha-1-Antitrypsin are shown.
[0473] Alpha-1-Antitrypsin is associated with alpha-1-Antitrypsin
deficiency disease, cystic fibrosis, interstitial lung disease, and
pulmonary arterial hypertension.
[0474] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the 5'-UTR of the
native mRNA of alpha-1-Antitrypsin. The complete mUNA molecule
comprises a 5' cap (m7GpppGm) upstream of the sequence below, and
coding region (CDS) for human alpha-1-Antitrypsin, a 3' UTR and
polyA tail (SEQ ID Nos: 4 to 12) downstream of the sequence below,
each of which corresponds to the structure of the native mRNA of
alpha-1-Antitrypsin.
TABLE-US-00010 Human alpha-1-antitrypsin mRNA is accession
NM_000295.4. (SEQ ID NO: 21)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 22)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCCGA{circumflex over
(C)}{circumflex over (C)} (SEQ ID NO: 23)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCC{circumflex over (G)}{tilde
over (A)}CC (SEQ ID NO: 24)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAG{circumflex over (C)}{circumflex
over (C)}GACC (SEQ ID NO: 25)
GGCACCACCACUGACCUGGGACAGUGAAUCGAC{tilde over (A)}{circumflex over
(G)}CCGACC (SEQ ID NO: 26) GGCACCACCACUGACCUGGGACAGUGAAUCG{tilde
over (A)}{circumflex over (C)}AGCCGACC (SEQ ID NO: 27)
GGCACCACCACUGACCUGGGACAGUGAAU{circumflex over (C)}{circumflex over
(G)}ACAGCCGACC (SEQ ID NO: 28) GGCACCACCACUGACCUGGGACAGUGA{tilde
over (A)}{tilde over (U)}CGACAGCCGACC (SEQ ID NO: 29)
GGCACCACCACUGACCUGGGACAGU{circumflex over (G)}{tilde over
(A)}AUCGACAGCCGACC (SEQ ID NO: 30)
GGCACCACCACUGACCUGGGACA{circumflex over (G)}{tilde over
(U)}GAAUCGACAGCCGACC (SEQ ID NO: 31)
GGCACCACCACUGACCUGGGA{circumflex over (C)}{tilde over
(A)}GUGAAUCGACAGCCGACC (SEQ ID NO: 32)
GGCACCACCACUGACCUGG{circumflex over (G)}{tilde over
(A)}CAGUGAAUCGACAGCCGACC (SEQ ID NO: 33)
GGCACCACCACUGACCU{circumflex over (G)}{circumflex over
(G)}GACAGUGAAUCGACAGCCGACC (SEQ ID NO: 34)
GGCACCACCACUGAC{circumflex over (C)}{tilde over
(U)}GGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 35) GGCACCACCACUG{tilde
over (A)}{circumflex over (C)}UGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO:
36) GGCACCACCAC{tilde over (U)}{circumflex over
(G)}ACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 37) GGCACCACC{tilde
over (A)}{circumflex over (C)}CUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ
ID NO: 38) GGCACCA{circumflex over (C)}{circumflex over
(C)}ACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 39)
GGCAC{circumflex over (C)}{tilde over
(A)}CCACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 40) GGC{tilde
over (A)}{circumflex over (C)}CACCACUGACCUGGGACAGUGAAUCGACAGCCGACC
(SEQ ID NO: 41) G{circumflex over (G)}{circumflex over
(C)}ACCACCACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 42)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCCG{tilde over (A)}C{circumflex
over (C)} (SEQ ID NO: 43)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCC{circumflex over
(G)}AC{circumflex over (C)} (SEQ ID NO: 44)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGC{circumflex over
(C)}AC{circumflex over (C)} (SEQ ID NO: 45)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAG{circumflex over
(C)}CGAC{circumflex over (C)} (SEQ ID NO: 46)
GGCACCACCACUGACCUGGGACAGUGAAUCGACA{circumflex over
(G)}CCGAC{circumflex over (C)} (SEQ ID NO: 47)
GGCACCACCACUGACCUGGGACAGUGAAUCGAC{tilde over (A)}GCCGAC{circumflex
over (C)} (SEQ ID NO: 48)
GGCACCACCACUGACCUGGGACAGUGAAUCGA{circumflex over
(C)}AGCCGAC{circumflex over (C)} (SEQ ID NO: 49)
GGCACCACCACUGACCUGGGACAGUGAAUCG{tilde over (A)}CAGCCGAC{circumflex
over (C)} (SEQ ID NO: 50) GGCACCACCACUGACCUGGGACAGUGAAUC{circumflex
over (G)}ACAGCCGAC{circumflex over (C)} (SEQ ID NO: 51)
GGCACCACCACUGACCUGGGACAGUGAAU{circumflex over
(C)}GACAGCCGAC{circumflex over (C)} (SEQ ID NO: 52)
GGCACCACCACUGACCUGGGACAGUGAA{tilde over (U)}CGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 53) GGCACCACCACUGACCUGGGACAGUGA{tilde over
(A)}UCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 54)
GGCACCACCACUGACCUGGGACAGUG{tilde over (A)}AUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 55) GGCACCACCACUGACCUGGGACAGU{circumflex over
(G)}AAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 56)
GGCACCACCACUGACCUGGGACAG{tilde over (U)}GAAUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 57) GGCACCACCACUGACCUGGGACA{circumflex over
(G)}UGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 58)
GGCACCACCACUGACCUGGGAC{tilde over (A)}GUGAAUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 59) GGCACCACCACUGACCUGGGA{circumflex over
(C)}AGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 60)
GGCACCACCACUGACCUGGG{tilde over (A)}CAGUGAAUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 61) GGCACCACCACUGACCUGG{circumflex over
(G)}ACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 62)
GGCACCACCACUGACCUG{circumflex over
(G)}GACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 63)
GGCACCACCACUGACCU{circumflex over
(G)}GGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 64)
GGCACCACCACUGACC{tilde over (U)}GGGACAGUGAAUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 65) GGCACCACCACUGAC{circumflex over
(C)}UGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 66)
GGCACCACCACUGA{circumflex over
(C)}UUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 67)
GGCACCACCACUG{tilde over (A)}CCUGGGACAGUGAAUCGACAGCCGAC{circumflex
over (C)} (SEQ ID NO: 68) GGCACCACCACU{circumflex over
(G)}ACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
69) GGCACCACCAC{tilde over
(U)}GACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
70) GGCACCACCA{circumflex over
(C)}UGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
71) GGCACCACC{tilde over
(A)}CUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
72) GGCACCAC{circumflex over
(C)}ACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 73) GGCACCA{circumflex over
(C)}CACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 74) GGCACC{tilde over
(A)}CCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 75) GGCAC{circumflex over
(C)}ACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 76) GGCA{circumflex over
(C)}CACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 77) GGC{tilde over
(A)}CCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 78) GG{circumflex over
(C)}ACCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 79) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)}
(SEQ ID NO: 80) GGCACCACCACUGACCUGGGACAGUGAAUCGACAGCCG{tilde over
(A)}{circumflex over (C)}{circumflex over (C)} (SEQ ID NO: 81)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAG{circumflex over (C)}{circumflex
over (C)}{circumflex over (G)}ACC (SEQ ID NO: 82)
GGCACCACCACUGACCUGGGACAGUGAAUCGA{circumflex over (C)}{tilde over
(A)}{circumflex over (G)}CCGACC (SEQ ID NO: 83)
GGCACCACCACUGACCUGGGACAGUGAAUCGACAGC{circumflex over
(C)}{circumflex over
(G)}{tilde over (A)}CC (SEQ ID NO: 84)
GGCACCACCACUGACCUGGGACAGUG{tilde over (A)}{tilde over (A)}{tilde
over (U)}CGACAGCCGACC (SEQ ID NO: 85)
GGCACCACCACUGACCUGGGACA{circumflex over (G)}{tilde over
(U)}{circumflex over (G)}AAUCGACAGCCGACC (SEQ ID NO: 86)
GGCACCACCACUGACCUGGG{tilde over (U)}{circumflex over (C)}{tilde
over (A)}GUGAAUCGACAGCCGACC (SEQ ID NO: 87)
GGCACCACCACUGACCU{circumflex over (G)}{circumflex over
(G)}{circumflex over (G)}ACAGUGAAUCGACAGCCGACC (SEQ ID NO: 88)
GGCACCACCACUGA{circumflex over (C)}{circumflex over (C)}{tilde over
(U)}GGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 89) GGCACCACCAC{tilde over
(U)}{circumflex over (G)}{tilde over
(A)}CCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 90) GGCACCAC{circumflex
over (C)}{tilde over (A)}{circumflex over
(C)}UGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 91) GGCAC{circumflex
over (C)}{tilde over (A)}{circumflex over
(C)}CACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 92) GG{circumflex
over (C)}{tilde over (A)}{circumflex over
(C)}CACCACUGACCUGGGACAGUGAAUCGACAGCCGACC (SEQ ID NO: 93)
G{circumflex over (G)}CACCACCACUGACCUGGGACAGUGAAUCGACAGCCG{tilde
over (A)}C{circumflex over (C)} (SEQ ID NO: 94) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACAGCC{circumflex over
(G)}AC{circumflex over (C)} (SEQ ID NO: 95) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACAGC{circumflex over
(C)}GAC{circumflex over (C)} (SEQ ID NO: 96) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACAG{circumflex over
(C)}CGAC{circumflex over (C)} (SEQ ID NO: 97) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACA{circumflex over
(G)}CCGAC{circumflex over (C)} (SEQ ID NO: 98) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCG{tilde over
(A)}CAGCCGAC{circumflex over (C)} (SEQ ID NO: 99) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGA{circumflex over
(C)}AGCCGAC{circumflex over (C)} (SEQ ID NO: 100) G{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCG{tilde over
(A)}CAGCCGAC{circumflex over (C)} (SEQ ID NO: 101) G{circumflex
over (G)}CACCACCACUGACCUGGGACAGUGAAUC{circumflex over
(G)}ACAGCCGAC{circumflex over (C)} (SEQ ID NO: 102) G{circumflex
over (G)}CACCACCACUGACCUGGGACAGUGAAU{circumflex over
(C)}GACAGCCGAC{circumflex over (C)} (SEQ ID NO: 103) G{circumflex
over (G)}CACCACCACUGACCUGGGACAGUGAA{tilde over
(U)}CGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 104) G{circumflex
over (G)}CACCACCACUGACCUGGGACAGUGA{tilde over
(A)}UCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 105) G{circumflex
over (G)}CACCACCACUGACCUGGGACAGUG{tilde over
(A)}AUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 106)
G{circumflex over (G)}CACCACCACUGACCUGGGACAGU{circumflex over
(G)}AAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 107)
G{circumflex over (G)}CACCACCACUGACCUGGGACAG{tilde over
(U)}GAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 108)
G{circumflex over (G)}CACCACCACUGACCUGGGACA{circumflex over
(G)}UGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 109)
G{circumflex over (G)}CACCACCACUGACCUGGGAC{tilde over
(A)}GUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 110)
G{circumflex over (G)}CACCACCACUGACCUGGGA{circumflex over
(C)}AGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 111)
G{circumflex over (G)}CACCACCACUGACCUGGG{tilde over
(A)}CAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 112)
G{circumflex over (G)}CACCACCACUGACCUGG{circumflex over
(G)}ACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 113)
G{circumflex over (G)}CACCACCACUGACCUG{circumflex over
(G)}GACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 114)
G{circumflex over (G)}CACCACCACUGACCU{circumflex over
(G)}GGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 115)
G{circumflex over (G)}CACCACCACUGACC{tilde over
(U)}GGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 116)
G{circumflex over (G)}CACCACCACUGAC{circumflex over
(C)}UGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 117)
G{circumflex over (G)}CACCACCACUGA{circumflex over
(C)}CUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO: 118)
G{circumflex over (G)}CACCACCACUG{tilde over
(A)}CCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
119) G{circumflex over (G)}CACCACCACU{circumflex over
(G)}ACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
120) G{circumflex over (G)}CACCACCAC{tilde over
(U)}GACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
121) G{circumflex over (G)}CACCACCA{circumflex over
(C)}UGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID NO:
122) G{circumflex over (G)}CACC{tilde over
(A)}CCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 123) G{circumflex over (G)}CACCAC{circumflex over
(C)}ACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 124) G{circumflex over (G)}CACCA{circumflex over
(C)}CACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 125) G{circumflex over (G)}CACC{tilde over
(A)}CCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 126) G{circumflex over (G)}CAC{circumflex over
(C)}ACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ ID
NO: 127) G{circumflex over (G)}CA{circumflex over
(C)}CACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 128) G{circumflex over (G)}C{tilde over
(A)}CCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 129) G{circumflex over (G)}{circumflex over
(C)}ACCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)} (SEQ
ID NO: 130) {circumflex over (G)}{circumflex over
(G)}CACCACCACUGACCUGGGACAGUGAAUCGACAGCCGAC{circumflex over (C)}
Example 16
mUNA Oligomer Expressing Erythropoietin (EPO)
[0475] In this example, the structures of mUNA molecules for use in
expressing human Erythropoietin (EPO) are shown.
[0476] Erythropoietin is available as a commercial drug and is
indicated for anemia resulting from chronic kidney disease,
inflammatory bowel disease including Crohn's disease and ulcer
colitis, and myelodysplasia from the treatment of cancer with
chemotherapy or radiation.
[0477] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human Erythropoietin. The complete mUNA
molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream of
the sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4 to
12) downstream of the sequence below, each of which corresponds to
the structure of the native mRNA of human Erythropoietin.
TABLE-US-00011 Human Erythropoietin is accession NM_000799.2. (SEQ
ID NO: 131) A{tilde over (U)}{circumflex over (G)}G{circumflex over
(G)}GGUGCACGAAUGUCCUGCCUGGCUGUGGCUUCUCCUGUCCCUGC
UGUCGCUCCCUCUGGGCCUCCCAGUCCUGGGCGCCCCACCACGCCUCAU
CUGUGACAGCCGAGUCCUGGAGAGGUACCUCUUGGAGGCCAAGGAGGCC
GAGAAUAUCACGACGGGCUGUGCUGAACACUGCAGCUUGAAUGAGAAUA
UCACUGUCCCAGACACCAAAGUUAAUUUCUAUGCCUGGAAGAGGAUGGA
GGUCGGGCAGCAGGCCGUAGAAGUCUGGCAGGGCCUGGCCCUGCUGUCG
GAAGCUGUCCUGCGGGGCCAGGCCCUGUUGGUCAACUCUUCCCAGCCGU
GGGAGCCCCUGCAGCUGCAUGUGGAUAAAGCCGUCAGUGGCCUUCGCAG
CCUCACCACUCUGCUUCGGGCUCUGGGAGCCCAGAAGGAAGCCAUCUCC
CCUCCAGAUGCGGCCUCAGCUGCUCCACUCCGAACAAUCACUGCUGACA
CUUUCCGCAAACUCUUCCGAGUCUACUCCAAUUUCCUCCGGGGAAAGCU
GAAGCUGUACACAGGGGAGGCCUGCAGGACAGGGGACAG{tilde over (A)}{tilde over
(U)}{circumflex over (G)}A (SEQ ID NO: 132) AU{circumflex over
(G)}GGGGUGCACGA{tilde over (A)}UGUCCUGCCUG{circumflex over
(G)}CUGUGGCUUCU{circumflex over (C)}CUGUCCCUGC U{circumflex over
(G)}UCGCUCCCUCU{circumflex over (G)}GGCCUCCCAGU{circumflex over
(C)}CUGGGCGCCCC{tilde over (A)}CCACGCCUCAU {circumflex over
(C)}UGUGACAGCCG{tilde over (A)}GUCCUGGAGAG{circumflex over
(G)}UACCUCUUGGA{circumflex over (G)}GCCAAGGAGGC {circumflex over
(C)}GAGAAUAUCAC{circumflex over (G)}ACGGGCUGUGC{tilde over
(U)}GAACACUGCAG{circumflex over (C)}UUGAAUGAGAA {tilde over
(U)}AUCACUGUCCC{tilde over (A)}GACACCAAAGU{tilde over
(U)}AAUUUCUAUGC{circumflex over (C)}UGGAAGAGGAU {circumflex over
(G)}GAGGUCGGGCA{circumflex over (G)}CAGGCCGUAGA{tilde over
(A)}GUCUGGCAGGG{circumflex over (C)}UUGGCCCUGCU {circumflex over
(G)}UCGGAAGCUGU{circumflex over (C)}CUGCGGGGCCA{circumflex over
(G)}GCCCUGUUGGU{circumflex over (C)}AACUCUUCCC A{circumflex over
(G)}CCGUGGGAGCC{circumflex over (G)}CUGCAGCUGCA{tilde over
(U)}GUGGAUAAAGC{circumflex over (C)}GUCAGUGGCC U{tilde over
(U)}CGCAGCCUCAC{circumflex over
(C)}ACUCUGCUUCGGGCUCUGGGAGC{circumflex over (C)}CAGAAGGAAG
CCAUCUCCCCUCC{tilde over (A)}GAUGCGGCCUC{tilde over
(A)}GCUGCUCCACU{circumflex over (C)}CGAACAAUCA C{tilde over
(U)}GCUGACACUUU{circumflex over (C)}CGCAAACUCUU{circumflex over
(C)}CGAGUCUACUC{circumflex over (C)}AAUUUCCUCC G{circumflex over
(G)}GGAAAGCUGAA{circumflex over (G)}CUGUACACAGG{circumflex over
(G)}GAGGCCUGCAG{circumflex over (G)}ACAGGGGAC AG{tilde over (A)}UGA
(SEQ ID NO: 133) A{tilde over (U)}GGGGG{tilde over
(U)}GCACGAA{tilde over (U)}G{tilde over (U)}CC{tilde over
(U)}GCC{tilde over (U)}GGC{tilde over (U)}G{tilde over
(U)}GGC{tilde over (U)}{tilde over (U)}C{tilde over (U)}CC{tilde
over (U)}G{tilde over (U)}CCC{tilde over (U)}GC {tilde over
(U)}G{tilde over (U)}CGC{tilde over (U)}CCC{tilde over (U)}C{tilde
over (U)}GGGCC{tilde over (U)}CCCAG{tilde over (U)}CC{tilde over
(U)}GGGCGCCCCACCACGCC{tilde over (U)}CA{tilde over (U)} C{tilde
over (U)}G{tilde over (U)}GACAGCCGAG{tilde over (U)}CC{tilde over
(U)}GGAGAGG{tilde over (U)}ACC{tilde over (U)}C{tilde over
(U)}{tilde over (U)}GGAGGCCAAGGAGGCC GAGAA{tilde over (U)}A{tilde
over (U)}CACGACGGGC{tilde over (U)}G{tilde over (U)}GC{tilde over
(U)}GAACAC{tilde over (U)}GCAGC{tilde over (U)}{tilde over
(U)}GAA{tilde over (U)}GAGAA{tilde over (U)}A {tilde over
(U)}CAC{tilde over (U)}G{tilde over (U)}CCCAGACACCAAAG{tilde over
(U)}{tilde over (U)}AA{tilde over (U)}{tilde over (U)}{tilde over
(U)}C{tilde over (U)}A{tilde over (U)}GCC{tilde over
(U)}GGAAGAGGA{tilde over (U)}GGA GG{tilde over
(U)}CGGGCAGCAGGCCG{tilde over (U)}AGAAG{tilde over (U)}C{tilde over
(U)}GGCAGGGCC{tilde over (U)}GGCCC{tilde over (U)}GC{tilde over
(U)}G{tilde over (U)}CG GAAGC{tilde over (U)}G{tilde over
(U)}CC{tilde over (U)}GCGGGGCCAGGCCC{tilde over (U)}G{tilde over
(U)}{tilde over (U)}GG{tilde over (U)}CAAC{tilde over (U)}C{tilde
over (U)}{tilde over (U)}CCCAGCCG{tilde over (U)} GGGAGCCCC{tilde
over (U)}GCAGC{tilde over (U)}GCA{tilde over (U)}G{tilde over
(U)}GGA{tilde over (U)}AAAGCCG{tilde over (U)}CAG{tilde over
(U)}GGCC{tilde over (U)}{tilde over (U)}CGCAG CC{tilde over
(U)}CACCAC{tilde over (U)}C{tilde over (U)}GC{tilde over (U)}{tilde
over (U)}CGGGC{tilde over (U)}C{tilde over
(U)}GGGAGCCCAGAAGGAAGCCA{tilde over (U)}C{tilde over (U)}CC
CC{tilde over (U)}CCAGA{tilde over (U)}GCGGCC{tilde over
(U)}CAGC{tilde over (U)}GC{tilde over (U)}CCAC{tilde over
(U)}CCGAACAA{tilde over (U)}CAC{tilde over (U)}GC{tilde over
(U)}GACA C{tilde over (U)}{tilde over (U)}{tilde over
(U)}CCGCAAAC{tilde over (U)}C{tilde over (U)}{tilde over
(U)}CCGAG{tilde over (U)}C{tilde over (U)}AC{tilde over
(U)}CCAA{tilde over (U)}{tilde over (U)}{tilde over (U)}CC{tilde
over (U)}CCGGGGAAAGC{tilde over (U)} GAAGC{tilde over (U)}G{tilde
over (U)}ACACAGGGGAGGCC{tilde over (U)}GCAGGACAGGGGACAGA{tilde over
(U)}GA
Example 17
mUNA Oligomer Expressing Ornithine Transcarbamylase
[0478] In this example, the structures of mUNA molecules for use in
expressing human Ornithine transcarbamylase are shown.
[0479] Ornithine transcarbamylase is associated with Ornithine
transcarbamylase deficiency.
[0480] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human Ornithine transcarbamylase. The
complete mUNA molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR
upstream of the sequence below, and a 3' UTR and polyA tail (SEQ ID
Nos: 4 to 12) downstream of the sequence below, each of which
corresponds to the structure of the native mRNA of human Ornithine
transcarbamylase.
TABLE-US-00012 Human Ornithine transcarbamylase is accession
NM_000531.5. (SEQ ID NO: 134) AU{circumflex over
(G)}CUGUUUAAUCU{circumflex over (G)}AGGAUCCUGUU{tilde over
(U)}AAACAAUGCAG{circumflex over (C)}UUUUAGAAAUG{circumflex over
(G)}UCACAACU UCA{tilde over (U)}GGUUCGAAAUU{tilde over
(U)}UCGGUGUGGAC{tilde over (A)}ACCACUACAAA{tilde over
(A)}UAAAGUGCAGC{tilde over (U)}GAAGGGC CGUG{tilde over
(A)}CCUUCUCACUC{tilde over (U)}AAAAAACUUUA{circumflex over
(C)}CGGAGAAGAAA{tilde over (U)}UAAAUAUAUGC{tilde over (U)}AUGGCU
AUCAG{circumflex over (C)}AGAUCUGAAAU{tilde over
(U)}UAGGAUAAAAC{tilde over (A)}GAAAGGAGAGU{tilde over
(A)}UUUGCCUUUAU{tilde over (U)}GCAAG GGAAGU{circumflex over
(C)}CUUAGGCAUGA{tilde over (U)}UUUUGAGAAAA{circumflex over
(G)}AAGUACUCGAA{circumflex over (C)}AAGAUUGUCUA{circumflex over
(C)}AGAA ACAGGCU{tilde over (U)}UGCACUUCUGG{circumflex over
(G)}AGGACAUCCUU{circumflex over (G)}UUUUCUUACCA{circumflex over
(C)}ACAAGAUAUUC{tilde over (A)}UUU GGGUGUGA{tilde over
(A)}UGAAAGUCUCA{circumflex over (C)}GGACACGGCCC{circumflex over
(G)}UGUAUUGUCUAGCAU{circumflex over (G)}GCAGAUG{circumflex over
(C)}AG UAUUGGCUC{circumflex over (G)}AGUGUAUAAAC{tilde over
(A)}AUCAGAUUUGG{tilde over (A)}CACCCUGGCUA{tilde over
(A)}AGAAGCAUCCA{tilde over (U)}C CCAAUUAUCA{tilde over
(A)}UGGGCUGUCAG{tilde over (A)}UUUGUACCAUC{circumflex over
(C)}UAUCCAGAUCC{tilde over (U)}GGCUGAUUACC{tilde over (U)}
CACGCUCCAGG{tilde over (A)}ACACUAUAGCU{circumflex over
(C)}UCUGAAAGGUC{tilde over (U)}UACCCUCAGCU{circumflex over
(G)}GAUCGGGGAUG {circumflex over (G)}GAACAAUAUCC{tilde over
(U)}GCACUCCAUCA{tilde over (U)}GAUGAGCGCAG{circumflex over
(C)}GAAAUUCGGAA{tilde over (U)}GCACCUUCAG G{circumflex over
(C)}AGCUACUCCAA{tilde over (A)}GGGUUAUGAGC{circumflex over
(C)}GGAUGCUAGUG{tilde over (U)}AACCAAGUUGG{circumflex over
(C)}AGAGCAGUA UG{circumflex over (C)}CAAAGAGAAUG{circumflex over
(G)}UACCAAGCUGU{tilde over (U)}GCUGACAAAUG{tilde over
(A)}UCCAUUGGAAG{circumflex over (C)}AGCGCAUG GAG{circumflex over
(G)}CAAUGUAUUAA{tilde over (U)}UACAGACACUU{circumflex over
(G)}GAUAAGCAUGG{circumflex over (G)}ACAAGAAGAGG{tilde over
(A)}GAAGAAA AAGC{circumflex over (G)}GCUCCAGGCUU{tilde over
(U)}CCAAGGUUACC{tilde over (A)}GGUUACAAUGA{tilde over
(A)}GACUGCUAAAG{tilde over (U)}UGCUGC CUCUG{tilde over
(A)}CUGGACAUUUU{tilde over (U)}ACACUGCUUGC{circumflex over
(C)}CAGAAAGCCAG{tilde over (A)}AGAAGUGGAUG{tilde over (A)}UGAAG
UCUUUU{tilde over (A)}UUCUCCUCGAU{circumflex over
(C)}ACUAGUGUUCC{circumflex over (C)}AGAGGCAGAAA{tilde over
(A)}CAGAAAGUGGA{circumflex over (C)}AAUC AUGGCUG{tilde over
(U)}CAUGGUGUCCC{tilde over (U)}GCUGACAGAUU{tilde over
(A)}CUCACCUCAGC{tilde over (U)}CCAGAAGCCUA{tilde over (A)}AUU
UU{circumflex over (G)}A (SEQ ID NO: 135) A{tilde over
(U)}{circumflex over (G)}{circumflex over
(C)}UGUUUAAUCUGAGGAUCCUGUUAAACAAUGCAGCUUUUAGAAAUGGUCACAACUU
CAUGGUUCGAAAUUUUCGGUGUGGACAACCACUACAAAAUAAAGUGCAGCUGAAGGGCC
GUGACCUUCUCACUCUAAAAAACUUUACCGGAGAAGAAAUUAAAUAUAUGCUAUGGCUA
UCAGCAGAUCUGAAAUUUAGGAUAAAACAGAAAGGAGAGUAUUUGCCUUUAUUGCAAGG
GAAGUCCUUAGGCAUGAUUUUUGAGAAAAGAAGUACUCGAACAAGAUUGUCUACAGAAA
CAGGCUUUGCACUUCUGGGAGGACAUCCUUGUUUUCUUACCACACAAGAUAUUCAUUUG
GGUGUGAAUGAAAGUCUCACGGACACGGCCCGUGUAUUGUCUAGCAUGGCAGAUGCAGU
AUUGGCUCGAGUGUAUAAACAAUCAGAUUUGGACACCCUGGCUAAAGAAGCAUCCAUCC
CAAUUAUCAAUGGGCUGUCAGAUUUGUACCAUCCUAUCCAGAUCCUGGCUGAUUACCUC
ACGCUCCAGGAACACUAUAGCUCUCUGAAAGGUCUUACCCUCAGCUGGAUCGGGGAUGG
GAACAAUAUCCUGCACUCCAUCAUGAUGAGCGCAGCGAAAUUCGGAAUGCACCUUCAGG
CAGCUACUCCAAAGGGUUAUGAGCCGGAUGCUAGUGUAACCAAGUUGGCAGAGCAGUAU
GCCAAAGAGAAUGGUACCAAGCUGUUGCUGACAAAUGAUCCAUUGGAAGCAGCGCAUGG
AGGCAAUGUAUUAAUUACAGACACUUGGAUAAGCAUGGGACAAGAAGAGGAGAAGAAAA
AGCGGCUCCAGGCUUUCCAAGGUUACCAGGUUACAAUGAAGACUGCUAAAGUUGCUGCC
UCUGACUGGACAUUUUUACACUGCUUGCCCAGAAAGCCAGAAGAAGUGGAUGAUGAAGU
CUUUUAUUCUCCUCGAUCACUAGUGUUCCCAGAGGCAGAAAACAGAAAGUGGACAAUCA
UGGCUGUCAUGGUGUCCCUGCUGACAGAUUACUCACCUCAGCUCCAGAAGCCUAAAUU{tilde
over (U)} {tilde over (U)}{circumflex over (G)}A (SEQ ID NO: 136)
A{tilde over (U)}GC{tilde over (U)}G{tilde over (U)}{tilde over
(U)}{tilde over (U)}AA{tilde over (U)}C{tilde over (U)}GAGGA{tilde
over (U)}CC{tilde over (U)}G{tilde over (U)}{tilde over
(U)}AAACAA{tilde over (U)}GCAGC{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}AGAAA{tilde over (U)}GG{tilde
over (U)}CACAAC{tilde over (U)}{tilde over (U)} CA{tilde over
(U)}GG{tilde over (U)}{tilde over (U)}CGAAA{tilde over (U)}{tilde
over (U)}{tilde over (U)}{tilde over (U)}CGG{tilde over (U)}G{tilde
over (U)}GGACAACCAC{tilde over (U)}ACAAAA{tilde over (U)}AAAG{tilde
over (U)}GCAGC{tilde over (U)}GAAGGGCC G{tilde over (U)}GACC{tilde
over (U)}{tilde over (U)}C{tilde over (U)}CAC{tilde over
(U)}C{tilde over (U)}AAAAAAC{tilde over (U)}{tilde over (U)}{tilde
over (U)}ACCGGAGAAGAAA{tilde over (U)}{tilde over (U)}AAA{tilde
over (U)}A{tilde over (U)}A{tilde over (U)}GC{tilde over
(U)}A{tilde over (U)}GGC{tilde over (U)}A {tilde over
(U)}CAGCAGA{tilde over (U)}C{tilde over (U)}GAAA{tilde over
(U)}{tilde over (U)}{tilde over (U)}AGGA{tilde over
(U)}AAAACAGAAAGGAGAG{tilde over (U)}A{tilde over (U)}{tilde over
(U)}{tilde over (U)}GCC{tilde over (U)}{tilde over (U)}{tilde over
(U)}A{tilde over (U)}{tilde over (U)}GCAAGG GAAG{tilde over
(U)}CC{tilde over (U)}{tilde over (U)}AGGCA{tilde over (U)}GA{tilde
over (U)}{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}GAGAAAAGAAG{tilde over (U)}AC{tilde over
(U)}CGAACAAGA{tilde over (U)}{tilde over (U)}G{tilde over
(U)}C{tilde over (U)}ACAGAAA CAGGC{tilde over (U)}{tilde over
(U)}{tilde over (U)}GCAC{tilde over (U)}{tilde over (U)}C{tilde
over (U)}GGGAGGACA{tilde over (U)}CC{tilde over (U)}{tilde over
(U)}G{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde over
(U)}C{tilde over (U)}{tilde over (U)}ACCACACAAGA{tilde over
(U)}A{tilde over (U)}{tilde over (U)}CA{tilde over (U)}{tilde over
(U)}{tilde over (U)}G GG{tilde over (U)}G{tilde over (U)}GAA{tilde
over (U)}GAAAG{tilde over (U)}C{tilde over
(U)}CACGGACACGGCCCG{tilde over (U)}G{tilde over (U)}A{tilde over
(U)}{tilde over (U)}G{tilde over (U)}C{tilde over (U)}AGCA{tilde
over (U)}GGCAGA{tilde over (U)}GCAG{tilde over (U)} A{tilde over
(U)}{tilde over (U)}GGC{tilde over (U)}CGAG{tilde over (U)}G{tilde
over (U)}A{tilde over (U)}AAACAA{tilde over (U)}CAGA{tilde over
(U)}{tilde over (U)}{tilde over (U)}GGACACCC{tilde over
(U)}GGC{tilde over (U)}AAAGAAGCA{tilde over (U)}CCA{tilde over
(U)}CC CAA{tilde over (U)}{tilde over (U)}A{tilde over
(U)}CAA{tilde over (U)}GGGC{tilde over (U)}G{tilde over
(U)}CAGA{tilde over (U)}{tilde over (U)}{tilde over (U)}G{tilde
over (U)}ACCA{tilde over (U)}CC{tilde over (U)}A{tilde over
(U)}CCAGA{tilde over (U)}CC{tilde over (U)}GGC{tilde over
(U)}GA{tilde over (U)}{tilde over (U)}ACC{tilde over (U)}C
ACGC{tilde over (U)}CCAGGAACAC{tilde over (U)}A{tilde over
(U)}AGC{tilde over (U)}C{tilde over (U)}C{tilde over
(U)}GAAAGG{tilde over (U)}C{tilde over (U)}{tilde over
(U)}ACCC{tilde over (U)}CAGC{tilde over (U)}GGA{tilde over
(U)}CGGGGA{tilde over (U)}GG GAACAA{tilde over (U)}A{tilde over
(U)}CC{tilde over (U)}GCAC{tilde over (U)}CCA{tilde over
(U)}CA{tilde over (U)}GA{tilde over (U)}GAGCGCAGCGAAA{tilde over
(U)}{tilde over (U)}CGGAA{tilde over (U)}GCACC{tilde over
(U)}{tilde over (U)}CAGG CAGC{tilde over (U)}AC{tilde over
(U)}CCAAAGGG{tilde over (U)}{tilde over (U)}A{tilde over
(U)}GAGCCGGA{tilde over (U)}GC{tilde over (U)}AG{tilde over
(U)}G{tilde over (U)}AACCAAG{tilde over (U)}{tilde over
(U)}GGCAGAGCAG{tilde over (U)}A{tilde over (U)} GCCAAAGAGAA{tilde
over (U)}GG{tilde over (U)}ACCAAGC{tilde over (U)}G{tilde over
(U)}{tilde over (U)}GC{tilde over (U)}GACAAA{tilde over
(U)}GA{tilde over (U)}CCA{tilde over (U)}{tilde over
(U)}GGAAGCAGCGCA{tilde over (U)}GG AGGCAA{tilde over (U)}G{tilde
over (U)}A{tilde over (U)}{tilde over (U)}AA{tilde over (U)}{tilde
over (U)}ACAGACAC{tilde over (U)}{tilde over (U)}GGA{tilde over
(U)}AAGCA{tilde over (U)}GGGACAAGAAGAGGAGAAGAAAA AGCGGC{tilde over
(U)}CCAGGC{tilde over (U)}{tilde over (U)}{tilde over
(U)}CCAAGG{tilde over (U)}{tilde over (U)}ACCAGG{tilde over
(U)}{tilde over (U)}ACAA{tilde over (U)}GAAGAC{tilde over
(U)}GC{tilde over (U)}AAAG{tilde over (U)}{tilde over (U)}GC{tilde
over (U)}GCC {tilde over (U)}C{tilde over (U)}GAC{tilde over
(U)}GGACA{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}ACAC{tilde over (U)}GC{tilde over
(U)}{tilde over (U)}GCCCAGAAAGCCAGAAGAAG{tilde over (U)}GGA{tilde
over (U)}GA{tilde over (U)}GAAG{tilde over (U)} C{tilde over
(U)}{tilde over (U)}{tilde over (U)}{tilde over (U)}A{tilde over
(U)}{tilde over (U)}C{tilde over (U)}CC{tilde over (U)}CGA{tilde
over (U)}CAC{tilde over (U)}AG{tilde over (U)}G{tilde over
(U)}{tilde over (U)}CCCAGAGGCAGAAAACAGAAAG{tilde over
(U)}GGACAA{tilde over (U)}CA {tilde over (U)}GGC{tilde over
(U)}G{tilde over (U)}CA{tilde over (U)}GG{tilde over (U)}G{tilde
over (U)}CCC{tilde over (U)}GC{tilde over (U)}GACAGA{tilde over
(U)}{tilde over (U)}AC{tilde over (U)}CACC{tilde over
(U)}CAGC{tilde over (U)}CCAGAAGCC{tilde over (U)}AAA{tilde over
(U)}{tilde over (U)}{tilde over (U)} {tilde over (U)}GA
Example 18
mUNA Oligomer Expressing Beta-Globin
[0481] In this example, the structures of mUNA molecules for use in
expressing human beta-globin are shown.
[0482] Beta-globin may be associated with sickle-cell disease, beta
thalassemia, and genetic resistance to malaria.
[0483] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the 3'-UTR of the
native mRNA of human beta-globin. The complete mUNA molecule
comprises a 5' cap (m7GpppGm), 5'-UTR, and coding region (CDS) for
human beta-globin upstream of the sequence below, and a polyA tail
(SEQ ID Nos: 4 to 12) downstream of the sequence below, each of
which corresponds to the structure of the native mRNA of human
beta-globin.
TABLE-US-00013 Human beta-globin is accession NM_000518.4. (SEQ ID
NO: 137) G{circumflex over
(C)}UCGCUUUCUUGCUGUCCAAUUUCUAUUAAAGGUUCCUUUGUUCCCUA
AGUCCAACUACUAAACUGGGGGAUAUUAUGAAGGGCCUUGAGCAUCUGG
AUUCUGCCUAAUAAAAAACAUUUAUUUUCAUUGC{tilde over (A)}A (SEQ ID NO:
138) G{circumflex over (C)}{tilde over (U)}{circumflex over
(C)}GCUUUCUUGCUGUCCAAUUUCUAUUAAAGGUUCCUUUGUUCCCUA
AGUCCAACUACUAAACUGGGGGAUAUUAUGAAGGGCCUUGAGCAUCUGG
AUUCUGCCUAAUAAAAAACAUUUAUUUUCAUU{circumflex over (G)}{circumflex
over (C)}{tilde over (A)}A (SEQ ID NO: 139) G{circumflex over
(C)}UCGCU{tilde over (U)}UCUUG{circumflex over (C)}UGUCC{tilde over
(A)}AUUUC{tilde over (U)}AUUAA{tilde over (A)}GGUUC{circumflex over
(C)}UUUGU{tilde over (U)}CCCUA {tilde over (A)}GUCCA{tilde over
(A)}CUACU{tilde over (A)}AACUG{circumflex over (G)}GGGAU{tilde over
(A)}UUAUG{tilde over (A)}AGGGC{circumflex over (C)}UUGAG{circumflex
over (C)}AUCUG{circumflex over (G)} AUUCU{circumflex over
(G)}CCUAA{tilde over (U)}AAAAA{tilde over (A)}CAUUU{tilde over
(A)}UUUUC{tilde over (A)}UUGC{tilde over (A)}A (SEQ ID NO: 140)
G{circumflex over (C)}{tilde over (U)}CGCU{tilde over (U)}{tilde
over (U)}CUUG{circumflex over (C)}{tilde over (U)}GUCC{tilde over
(A)}{tilde over (A)}UUUC{tilde over (U)}{tilde over (A)}UUAA{tilde
over (A)}{circumflex over (G)}GUUC{circumflex over (C)}{tilde over
(U)}UUGU{tilde over (U)}{circumflex over (C)}CCU A{tilde over
(A)}{circumflex over (G)}UCCA{tilde over (A)}{circumflex over
(C)}UACU{tilde over (A)}{tilde over (A)}ACUG{circumflex over
(G)}{circumflex over (G)}GGAU{tilde over (A)}{tilde over
(U)}UAUG{tilde over (A)}{tilde over (A)}GGGC{circumflex over
(C)}{tilde over (U)}UGAG{circumflex over (C)}{tilde over (A)}UCU
G{circumflex over (G)}{tilde over (A)}UUCU{circumflex over
(G)}{circumflex over (C)}CUAA{tilde over (U)}{tilde over
(A)}AAAA{tilde over (A)}{circumflex over (C)}AUUU{tilde over
(A)}{tilde over (U)}UUUC{tilde over (A)}{tilde over (U)}UGC{tilde
over (A)}A (SEQ ID NO: 141) {circumflex over (G)}{circumflex over
(C)}{tilde over (U)}{circumflex over (C)}{circumflex over
(G)}{circumflex over (C)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}{circumflex over (C)}{tilde over (U)}{tilde over
(U)}{circumflex over (G)}{circumflex over (C)}{tilde over
(U)}{circumflex over (G)}{tilde over (U)}{circumflex over
(C)}{circumflex over (C)}{tilde over (A)}{tilde over (A)}{tilde
over (U)}{tilde over (U)}{tilde over (U)}{circumflex over
(C)}{tilde over (U)}{tilde over (A)}{tilde over (U)}{tilde over
(U)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{circumflex
over (G)}{circumflex over (G)}{tilde over (U)}{tilde over
(U)}{circumflex over (C)}{circumflex over (C)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}{circumflex over (G)}{tilde
over (U)}{tilde over (U)}{circumflex over (C)}{circumflex over
(C)}{circumflex over (C)}{tilde over (U)} {tilde over (A)}{tilde
over (A)}{circumflex over (G)}{tilde over (U)}{circumflex over
(C)}{circumflex over (C)}{tilde over (A)}{tilde over
(A)}{circumflex over (G)}{tilde over (U)}{tilde over
(A)}{circumflex over (C)}{tilde over (U)}{tilde over (A)}{tilde
over (A)}{tilde over (A)}{circumflex over (C)}{tilde over
(U)}{circumflex over (G)}{circumflex over (G)}{circumflex over
(G)}{circumflex over (G)}{circumflex over (G)}{tilde over
(A)}{tilde over (U)}{tilde over (A)}{tilde over (U)}{tilde over
(U)}{tilde over (A)}{tilde over (U)}{circumflex over (G)}{tilde
over (A)}{tilde over (A)}{circumflex over (G)}{circumflex over
(G)}{circumflex over (G)}{circumflex over (C)}{circumflex over
(C)}{tilde over (U)}{tilde over (U)}{circumflex over (G)}{tilde
over (A)}{circumflex over (G)}{circumflex over (C)}{tilde over
(A)}{tilde over (U)}{circumflex over (C)}{tilde over (U)}
{circumflex over (G)}{circumflex over (G)}{tilde over (A)}{tilde
over (U)}{tilde over (U)}{circumflex over (C)}{tilde over
(U)}{circumflex over (G)}{circumflex over (C)}{circumflex over
(C)}{tilde over (A)}{tilde over (U)}{tilde over (A)}{tilde over
(A)}{tilde over (U)}{tilde over (A)}{tilde over (A)}{tilde over
(A)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{circumflex
over (C)}{tilde over (A)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (A)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}{circumflex over (C)}{tilde over
(A)}{tilde over (U)}{tilde over (U)}{circumflex over
(G)}{circumflex over (C)}{tilde over (A)}{tilde over (A)}
Example 19
mUNA Oligomer Translation Enhancer Based on Xenopus Beta-Globin
3'UTR
[0484] In this example, the structures of mUNA molecules for use in
enhancing translational efficiency are shown.
[0485] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the 3'-UTR of
Xenopus beta-globin. The complete mUNA molecule comprises a 5' cap
(m7GpppGm), 5'-UTR, and coding region (CDS) upstream of the
sequence below, and a polyA tail (SEQ ID Nos: 4 to 12) downstream
of the sequence below, each of which corresponds to the structure
of a native human mRNA. Thus, a UNA oligomer incorprating the
oligomer fragment below can have enhanced translational
efficiency.
TABLE-US-00014 Xenopus beta-globin is accession NM_001096347.1.
(SEQ ID NO: 142) C{tilde over
(U)}AGUGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACC
CGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUU
GUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUU CUUCAC{tilde
over (A)}U (SEQ ID NO: 143) C{tilde over (U)}{tilde over
(A)}{circumflex over
(G)}UGACUGACUAGGAUCUGGUUACCACUAAACCAGCCUCAAGAACACC
CGAAUGGAGUCUCUAAGCUACAUAAUACCAACUUACACUUACAAAAUGUU
GUCCCCCAAAAUGUAGCCAUUCGUAUCUGCUCCUAAUAAAAAGAAAGUUU CUUC{tilde over
(A)}{circumflex over (C)}{tilde over (A)}U (SEQ ID NO: 144) C{tilde
over (U)}AGUGA{circumflex over (C)}UGACU{tilde over (A)}GGAUC{tilde
over (U)}GGUUA{circumflex over (C)}CACUA{tilde over
(A)}ACCAG{circumflex over (C)}CUCAA{circumflex over (G)}AAAC
{circumflex over (C)}CAAAU{circumflex over (G)}GAGUC{tilde over
(U)}CUAAG{circumflex over (C)}UACAU{tilde over (A)}AUACC{tilde over
(A)}ACUUA{circumflex over (C)}ACUUA{circumflex over
(C)}AAAAU{circumflex over (G)}UU GUC{circumflex over
(C)}CCCAA{tilde over (A)}AUGUA{circumflex over (G)}CCAUU{circumflex
over (C)}GUAUC{tilde over (U)}GCUCC{tilde over (U)}AAUAA{tilde over
(A)}AAGAA{tilde over (A)}GUUU C{tilde over (U)}UCAC{tilde over
(A)}U (SEQ ID NO: 145) C{tilde over (U)}{tilde over
(A)}GUGA{circumflex over (C)}{tilde over (U)}GACU{tilde over
(A)}{circumflex over (G)}GAUC{tilde over (U)}{circumflex over
(G)}GUUA{circumflex over (C)}{circumflex over (C)}ACUA{tilde over
(A)}{tilde over (A)}CCAG{circumflex over (C)}{circumflex over
(C)}UCAA{circumflex over (G)}{tilde over (A)}ACAC {circumflex over
(C)}{circumflex over (C)}GAAU{circumflex over (G)}{circumflex over
(G)}AGUC{tilde over (U)}{circumflex over (C)}UAAG{circumflex over
(C)}{tilde over (U)}ACAU{tilde over (A)}{tilde over (A)}UACC{tilde
over (A)}{tilde over (A)}CUUA{circumflex over (C)}{tilde over
(A)}CUUA{circumflex over (C)}{tilde over (A)}AAAU{circumflex over
(G)}{tilde over (U)}U GUC{circumflex over (C)}{circumflex over
(C)}CCAA{tilde over (A)}{tilde over (A)}UGUA{circumflex over
(G)}{circumflex over (C)}CAUU{circumflex over (C)}{circumflex over
(G)}UAUC{tilde over (U)}{circumflex over (G)}CUCC{tilde over
(U)}{tilde over (A)}AUAA{tilde over (A)}{tilde over (A)}AGAA{tilde
over (A)}{circumflex over (G)}UUU C{tilde over (U)}{tilde over
(U)}CAC{tilde over (A)}U (SEQ ID NO: 146) {circumflex over
(C)}{tilde over (U)}{tilde over (A)}{circumflex over (G)}{tilde
over (U)}{circumflex over (G)}{tilde over (A)}{circumflex over
(C)}{tilde over (U)}{circumflex over (G)}{tilde over
(A)}{circumflex over (C)}{tilde over (U)}{tilde over
(A)}{circumflex over (G)}{circumflex over (G)}{tilde over
(A)}{tilde over (U)}{circumflex over (C)}{tilde over
(U)}{circumflex over (G)}{circumflex over (G)}{tilde over
(U)}{tilde over (U)}{tilde over (A)}{circumflex over
(C)}{circumflex over (C)}{tilde over (A)}{circumflex over
(C)}{tilde over (U)}{tilde over (A)}{tilde over (A)}{tilde over
(A)}{circumflex over (C)}{circumflex over (C)}{tilde over
(A)}{circumflex over (G)}{circumflex over (C)}{circumflex over
(C)}{tilde over (U)}{circumflex over (C)}{tilde over (A)}{tilde
over (A)}{circumflex over (G)}{tilde over (A)}{tilde over
(A)}{circumflex over (C)}{tilde over (A)}{circumflex over (C)}
{circumflex over (C)}{circumflex over (C)}{tilde over (A)}{tilde
over (A)}{tilde over (A)}{tilde over (U)}{circumflex over
(G)}{circumflex over (G)}{tilde over (A)}{circumflex over
(G)}{tilde over (U)}{circumflex over (C)}{tilde over
(U)}{circumflex over (C)}{tilde over (U)}{tilde over (A)}{tilde
over (A)}{circumflex over (G)}{circumflex over (C)}{tilde over
(U)}{tilde over (A)}{circumflex over (C)}{tilde over (A)}{tilde
over (U)}{tilde over (A)}{tilde over (A)}{tilde over (U)}{tilde
over (A)}{circumflex over (C)}{circumflex over (C)}{tilde over
(A)}{tilde over (A)}{circumflex over (C)}{tilde over (U)}{tilde
over (U)}{tilde over (A)}{circumflex over (C)}{tilde over
(A)}{circumflex over (C)}{tilde over (U)}{tilde over (U)}{tilde
over (A)}{circumflex over (C)}{tilde over (A)}{tilde over
(A)}{tilde over (A)}{tilde over (A)}{tilde over (U)}{circumflex
over (G)}{tilde over (U)}{tilde over (U)} {circumflex over
(G)}{tilde over (U)}{circumflex over (C)}{circumflex over
(C)}{circumflex over (C)}{circumflex over (C)}{circumflex over
(C)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{tilde over
(A)}{tilde over (U)}{circumflex over (G)}{tilde over (U)}{tilde
over (A)}{circumflex over (G)}{circumflex over (C)}{circumflex over
(C)}{tilde over (A)}{tilde over (U)}{tilde over (U)}{circumflex
over (C)}{circumflex over (G)}{tilde over (U)}{tilde over
(A)}{tilde over (U)}{circumflex over (C)}{tilde over
(U)}{circumflex over (G)}{circumflex over (C)}{tilde over
(U)}{circumflex over (C)}{circumflex over (C)}{tilde over
(U)}{tilde over (A)}{tilde over (A)}{tilde over (U)}{tilde over
(A)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{tilde over
(A)}{circumflex over (G)}{tilde over (A)}{tilde over (A)}{tilde
over (A)}{circumflex over (G)}{tilde over (U)}{tilde over
(U)}{tilde over (U)} {circumflex over (C)}{tilde over (U)}{tilde
over (U)}{circumflex over (C)}{tilde over (A)}{circumflex over
(C)}{tilde over (A)}{tilde over (U)}
Example 20
mUNA Oligomer Expressing Thrombopoietin
[0486] In this example, the structures of mUNA molecules for use in
expressing human Thrombopoietin are shown.
[0487] Thrombopoietin is associated with liver and kidney
disease.
[0488] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human Thrombopoietin. The complete mUNA
molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream of
the sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4 to
12) downstream of the sequence below, each of which corresponds to
the structure of the native mRNA of human Thrombopoietin.
TABLE-US-00015 Human Thrombopoietin is accession NM_000460.3. (SEQ
ID NO: 147) AU{circumflex over (G)}GAGCUGACUGAAUUGCU{circumflex
over (C)}CUCGUGGUCAUGCUUCU{circumflex over (C)}CUAACUGCAA
GGCUAAC{circumflex over (G)}CUGUCCAGCCCGGCUCC{tilde over
(U)}CCUGCUUGUGACCUCCG{tilde over (A)}GUCCU CAGUAAACUGCU{tilde over
(U)}CGUGACUCCCAUGUCCU{tilde over (U)}CACAGCAGACUGAGCCA{circumflex
over (G)} UGCCCAG AGGUUCACCC{tilde over
(U)}UUGCCUACACCUGUCCU{circumflex over (G)}CUGCCUGCUGUGG ACUU{tilde
over (U)}AGCUUGGGAGAAUGGA{tilde over
(A)}AACCCAGAUGGAGGAGA{circumflex over (C)}CAAGGCACA GGACAUUC{tilde
over (U)}GGGAGCAGUGACCCUUC{tilde over
(U)}GCUGGAGGGAGUGAUGG{circumflex over (C)}AGCA
CGGGGACAACUGG{circumflex over (G)}ACCCACUUGCCUCUCAUC{circumflex
over (C)}CUCCUGGGGCAGCUUU C{tilde over
(U)}GGACAGGUCCGUCUCCU{circumflex over
(C)}CUUGGGGCCCUGCAGAG{circumflex over (C)}CUCCUUGGAAC CCAGCU{tilde
over (U)}CCUCCACAGGGCAGGAC{circumflex over
(C)}ACAGCUCACAAGGAUCCCAAUGCC AUCUUCCUGAG{circumflex over
(C)}UUCCAACACCUGCUCCG{tilde over (A)}GGAAAGGUGCGUUUCCUG{tilde over
(A)} UGCUUGUAGGAGGGUCC{tilde over (A)}CCCUCUGCGUCAGGCGGG{circumflex
over (G)}CCCCACCCACCA CAGCU{circumflex over
(G)}UCCCCAGCAGAACCUCU{circumflex over
(C)}UAGUCCUCACACUGAAC{circumflex over (G)}AGCUCCC AAACAGGACU{tilde
over (U)}CUGGAUUGUUGGAGACA{tilde over (A)}ACUUCACUGCCUCAGCC{tilde
over (A)}GA ACUACUGGCUCUGGG{circumflex over
(C)}UUCUGAAGUGGCAGCAG{circumflex over (G)}GAUUCAGAGCCAAGA
UU{circumflex over (C)}CUGGUCUGCUGAACCAA{tilde over
(A)}CCUCCAGGUCCCUGGAC{circumflex over (C)}AAAUCCCCGG AUACCUG{tilde
over (A)}ACAGGAUACACGAACUC{tilde over
(U)}UGAAUGGAACUCGUGGA{circumflex over (C)}UCUUU
CCUGGACCCUCA{circumflex over (C)}GCAGGACCCUAGGAGCC{circumflex over
(C)}CGGACAUUUCCUCAGGA{tilde over (A)} CAUCAGACACAGGCUCC{circumflex
over (C)}UGCCACCCAACCUCCAG{circumflex over (C)}CUGGAUAUUCUCC
UUCC{circumflex over (C)}CAACCCAUCCUCCUACU{circumflex over
(G)}GACAGUAUACGCUCUUC{circumflex over (C)}CUCUUCCA
CCCACCUUG{circumflex over (C)}CCACCCCUGUGGUCCAG{circumflex over
(C)}UCCACCCCCUGCUUCCU{circumflex over (G)}ACC
CUUCUGCUCCAACG{circumflex over (C)}CCACCCCUACCAGCCCU{circumflex
over (C)}UUCUAAACACAUCCUA C{tilde over (A)}CCCACUCCCAGAAUCUG{tilde
over (U)}CUCAGGAAGGGU{tilde over (A)}A (SEQ ID NO: 148) A{tilde
over (U)}{circumflex over (G)}{circumflex over
(G)}AGCUGACUGAAUUGCUCCUCGUGGUCAUGCUUCUCCUAACUGCAA
GGCUAACGCUGUCCAGCCCGGCUCCUCCUGCUUGUGACCUCCGAGUCCU
CAGUAAACUGCUUCGUGACUCCCAUGUCCUUCACAGCAGACUGAGCCAG
UGCCCAGAGGUUCACCCUUUGCCUACACCUGUCCUGCUGCCUGCUGUGG
ACUUUAGCUUGGGAGAAUGGAAAACCCAGAUGGAGGAGACCAAGGCACA
GGACAUUCUGGGAGCAGUGACCCUUCUGCUGGAGGGAGUGAUGGCAGCA
CGGGGACAACUGGGACCCACUUGCCUCUCAUCCCUCCUGGGGCAGCUUU
CUGGACAGGUCCGUCUCCUCCUUGGGGCCCUGCAGAGCCUCCUUGGAAC
CCAGCUUCCUCCACAGGGCAGGACCACAGCUCACAAGGAUCCCAAUGCC
AUCUUCCUGAGCUUCCAACACCUGCUCCGAGGAAAGGUGCGUUUCCUGA
UGCUUGUAGGAGGGUCCACCCUCUGCGUCAGGCGGGCCCCACCCACCAC
AGCUGUCCCCAGCAGAACCUCUCUAGUCCUCACACUGAACGAGCUCCCA
AACAGGACUUCUGGAUUGUUGGAGACAAACUUCACUGCCUCAGCCAGAA
CUACUGGCUCUGGGCUUCUGAAGUGGCAGCAGGGAUUCAGAGCCAAGAU
UCCUGGUCUGCUGAACCAAACCUCCAGGUCCCUGGACCAAAUCCCCGGA
UACCUGAACAGGAUACACGAACUCUUGAAUGGAACUCGUGGACUCUUUC
CUGGACCCUCACGCAGGACCCUAGGAGCCCCGGACAUUUCCUCAGGAAC
AUCAGACACAGGCUCCCUGCCACCCAACCUCCAGCCUGGAUAUUCUCCU
UCCCCAACCCAUCCUCCUACUGGACAGUAUACGCUCUUCCCUCUUCCAC
CCACCUUGCCCACCCCUGUGGUCCAGCUCCACCCCCUGCUUCCUGACCC
UUCUGCUCCAACGCCCACCCCUACCAGCCCUCUUCUAAACACAUCCUAC
ACCCACUCCCAGAAUCUGUCUCAGGAAGG{circumflex over (G)}{tilde over
(U)}{tilde over (A)}A (SEQ ID NO: 149) A{tilde over (U)}GGAGC{tilde
over (U)}GAC{tilde over (U)}GAA{tilde over (U)}{tilde over
(U)}GC{tilde over (U)}CC{tilde over (U)}CG{tilde over (U)}GG{tilde
over (U)}CA{tilde over (U)}GC{tilde over (U)}{tilde over
(U)}C{tilde over (U)}CC{tilde over (U)}AAC{tilde over (U)}GC
AAGGC{tilde over (U)}AACGC{tilde over (U)}G{tilde over
(U)}CCAGCCCGGC{tilde over (U)}CC{tilde over (U)}CC{tilde over
(U)}GC{tilde over (U)}G{tilde over (U)}GACC{tilde over (U)}CCGA
G{tilde over (U)}CC{tilde over (U)}CAG{tilde over (U)}AAAC{tilde
over (U)}GC{tilde over (U)}{tilde over (U)}CG{tilde over
(U)}GAC{tilde over (U)}CCCA{tilde over (U)}GCC{tilde over
(U)}{tilde over (U)}CACAGCAGA C{tilde over (U)}GAGCCAG{tilde over
(U)}GCCCAGAGG{tilde over (U)}{tilde over (U)}CACCC{tilde over
(U)}{tilde over (U)}{tilde over (U)}GCC{tilde over (U)}ACACC{tilde
over (U)}G{tilde over (U)}CC{tilde over (U)}G C{tilde over
(U)}GCC{tilde over (U)}GC{tilde over (U)}G{tilde over
(U)}GGAC{tilde over (U)}{tilde over (U)}{tilde over (U)}AGC{tilde
over (U)}{tilde over (U)}GGGAGAA{tilde over (U)}GGAAAACCCAGA{tilde
over (U)}G GAGGAGACCAAGGCACAGGACA{tilde over (U)}{tilde over
(U)}C{tilde over (U)}GGGAGCAG{tilde over (U)}GACCC{tilde over
(U)}C{tilde over (U)}GC{tilde over (U)} GGAGGGAG{tilde over
(U)}GA{tilde over (U)}GGCAGCACGGGGACAAC{tilde over
(U)}GGGACCCAC{tilde over (U)}{tilde over (U)}GCC{tilde over
(U)}C{tilde over (U)} CA{tilde over (U)}CCC{tilde over (U)}CC{tilde
over (U)}GGGGCAGC{tilde over (U)}{tilde over (U)}{tilde over
(U)}C{tilde over (U)}GGACAGG{tilde over (U)}CCG{tilde over
(U)}C{tilde over (U)}CC{tilde over (U)}CC{tilde over (U)}{tilde
over (U)}GG GGCCC{tilde over (U)}GCAGAGCC{tilde over (U)}CC{tilde
over (U)}{tilde over (U)}GGAACCCAGC{tilde over (U)}{tilde over
(U)}CC{tilde over (U)}CCACAGGGCAGGA CCACAGC{tilde over
(U)}CACAAGGA{tilde over (U)}CCCAA{tilde over (U)}GCCA{tilde over
(U)}C{tilde over (U)}{tilde over (U)}CC{tilde over (U)}GAGC{tilde
over (U)}{tilde over (U)}CCAACAC C{tilde over (U)}GC{tilde over
(U)}CCGAGGAAAGG{tilde over (U)}GCG{tilde over (U)}{tilde over
(U)}{tilde over (U)}CC{tilde over (U)}GA{tilde over (U)}GC{tilde
over (U)}{tilde over (U)}G{tilde over (U)}AGGAGGG{tilde over
(U)}CCAC CC{tilde over (U)}C{tilde over (U)}GCG{tilde over
(U)}CAGGCGGGCCCCACCCACCACAGC{tilde over (U)}G{tilde over
(U)}CCCCAGCAGAAC C{tilde over (U)}C{tilde over (U)}C{tilde over
(U)}AG{tilde over (U)}CC{tilde over (U)}CACAC{tilde over
(U)}GAACGAGC{tilde over (U)}CCCAAACAGGAC{tilde over (U)}{tilde over
(U)}C{tilde over (U)}GG A{tilde over (U)}{tilde over (U)}G{tilde
over (U)}{tilde over (U)}GGAGACAAAC{tilde over (U)}{tilde over
(U)}CAC{tilde over (U)}GCC{tilde over (U)}CAGCCAGAAC{tilde over
(U)}AC{tilde over (U)}GGC{tilde over (U)}C{tilde over (U)}G
GGC{tilde over (U)}C{tilde over (U)}GAAG{tilde over
(U)}GGCAGCAGGGA{tilde over (U)}{tilde over (U)}CAGAGCCAAGA{tilde
over (U)}{tilde over (U)}CC{tilde over (U)}GG{tilde over
(U)}C{tilde over (U)}G C{tilde over (U)}GAACCAAACC{tilde over
(U)}CCAGG{tilde over (U)}CCC{tilde over (U)}GGACCAAA{tilde over
(U)}CCCCGGA{tilde over (U)}ACC{tilde over (U)}GAACA GGA{tilde over
(U)}ACACGAAC{tilde over (U)}C{tilde over (U)}{tilde over
(U)}GAA{tilde over (U)}GGAAC{tilde over (U)}CG{tilde over
(U)}GGAC{tilde over (U)}C{tilde over (U)}{tilde over (U)}{tilde
over (U)}CC{tilde over (U)}GGACC C{tilde over
(U)}CACGCAGGACCC{tilde over (U)}AGGAGCCCCGGACA{tilde over
(U)}{tilde over (U)}{tilde over (U)}CC{tilde over
(U)}CAGGAACA{tilde over (U)}CAG ACACAGGC{tilde over (U)}CCC{tilde
over (U)}GCCACCCAACC{tilde over (U)}CCAGCC{tilde over (U)}GGA{tilde
over (U)}A{tilde over (U)}{tilde over (U)}C{tilde over (U)}CC{tilde
over (U)}{tilde over (U)}CC CCAACCCA{tilde over (U)}CC{tilde over
(U)}CC{tilde over (U)}AC{tilde over (U)}GGACAG{tilde over
(U)}A{tilde over (U)}ACGC{tilde over (U)}C{tilde over (U)}{tilde
over (U)}CCC{tilde over (U)}C{tilde over (U)}{tilde over (U)}CCACCC
ACC{tilde over (U)}{tilde over (U)}GCCCACCCC{tilde over (U)}G{tilde
over (U)}GG{tilde over (U)}CCAGC{tilde over (U)}CCACCCCC{tilde over
(U)}GC{tilde over (U)}{tilde over (U)}CC{tilde over (U)}GACCC
{tilde over (U)}{tilde over (U)}C{tilde over (U)}GC{tilde over
(U)}CCAACGCCCACCCC{tilde over (U)}ACCAGCCC{tilde over (U)}C{tilde
over (U)}{tilde over (U)}C{tilde over (U)}AAACACA{tilde over
(U)}CC{tilde over (U)} ACACCCAC{tilde over (U)}CCCAGAA{tilde over
(U)}C{tilde over (U)}G{tilde over (U)}C{tilde over
(U)}CAGGAAGGG{tilde over (U)}AA
Example 21
mUNA Oligomer Expressing Human Amylo-Alpha-1, 6-Glucosidase,
4-Alpha-Glucanotransferase (AGL)
[0489] In this example, the structures of mUNA molecules for use in
expressing human amylo-alpha-1, 6-glucosidase,
4-alpha-glucanotransferase (AGL) are shown.
[0490] AGL is associated with glycogen storage disease.
[0491] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human AGL. The complete mUNA molecule
comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream of the
sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4 to 12)
downstream of the sequence below, each of which corresponds to the
structure of the native mRNA of human AGL.
TABLE-US-00016 Human AGL is accession NM_000642.2. (SEQ ID NO: 150)
A{tilde over (U)}{circumflex over (G)}{circumflex over
(G)}GACACAGUAAACAGAUUCGAAUUUUACUUCUGAACGAAAUGGAGA
AACUGGAAAAGACCCUCUUCAGACUUGAACAAGGGUAUGAGCUACAGUU
CCGAUUAGGCCCAACUUUACAGGGAAAAGCAGUUACCGUGUAUACAAAU
UACCCAUUUCCUGGAGAAACAUUUAAUAGAGAAAAAUUCCGUUCUCUGG
AUUGGGAAAAUCCAACAGAAAGAGAAGAUGAUUCUGAUAAAUACUGUAA
ACUUAAUCUGCAACAAUCUGGUUCAUUUCAGUAUUAUUUCCUUCAAGGA
AAUGAGAAAAGUGGUGGAGGUUACAUAGUUGUGGACCCCAUUUUACGUG
UUGGUGCUGAUAAUCAUGUGCUACCCUUGGACUGUGUUACUCUUCAGAC
AUUUUUAGCUAAGUGUUUGGGACCUUUUGAUGAAUGGGAAAGCAGACUU
AGGGUUGCAAAAGAAUCAGGCUACAACAUGAUUCAUUUUACCCCAUUGC
AGACUCUUGGACUAUCUAGGUCAUGCUACUCCCUUGCCAAUCAGUUAGA
AUUAAAUCCUGACUUUUCAAGACCUAAUAGAAAGUAUACCUGGAAUGAU
GUUGGACAGCUAGUGGAAAAAUUAAAAAAGGAAUGGAAUGUUAUUUGUA
UUACUGAUGUUGUCUACAAUCAUACUGCUGCUAAUAGUAAAUGGAUCCA
GGAACAUCCAGAAUGUGCCUAUAAUCUUGUGAAUUCUCCACACUUAAAA
CCUGCCUGGGUCUUAGACAGAGCACUUUGGCGUUUCUCCUGUGAUGUUG
CAGAAGGGAAAUACAAAGAAAAGGGAAUACCUGCUUUGAUUGAAAAUGA
UCACCAUAUGAAUUCCAUCCGAAAAAUAAUUUGGGAGGAUAUUUUUCCA
AAGCUUAAACUCUGGGAAUUUUUCCAAGUAGAUGUCAACAAAGCGGUUG
AGCAAUUUAGAAGACUUCUUACACAAGAAAAUAGGCGAGUAACCAAGUC
UGAUCCAAACCAACACCUUACGAUUAUUCAAGAUCCUGAAUACAGACGG
UUUGGCUGUACUGUAGAUAUGAACAUUGCACUAACGACUUUCAUACCAC
AUGACAAGGGGCCAGCAGCAAUUGAAGAAUGCUGUAAUUGGUUUCAUAA
AAGAAUGGAGGAAUUAAAUUCAGAGAAGCAUCGACUCAUUAACUAUCAU
CAGGAACAGGCAGUUAAUUGCCUUUUGGGAAAUGUGUUUUAUGAACGAC
UGGCUGGCCAUGGUCCAAAACUAGGACCUGUCACUAGAAAGCAUCCUUU
AGUUACCAGGUAUUUUACUUUCCCAUUUGAAGAGAUAGACUUCUCCAUG
GAAGAAUCUAUGAUUCAUCUGCCAAAUAAAGCUUGUUUUCUGAUGGCAC
ACAAUGGAUGGGUAAUGGGAGAUGAUCCUCUUCGAAACUUUGCUGAACC
GGGUUCAGAAGUUUACCUAAGGAGAGAACUUAUUUGCUGGGGAGACAGU
GUUAAAUUACGCUAUGGGAAUAAACCAGAGGACUGUCCUUAUCUCUGGG
CACACAUGAAAAAAUACACUGAAAUAACUGCAACUUAUUUCCAGGGAGU
ACGUCUUGAUAACUGCCACUCAACACCUCUUCACGUAGCUGAGUACAUG
UUGGAUGCUGCUAGGAAUUUGCAACCCAAUUUAUAUGUAGUAGCUGAAC
UGUUCACAGGAAGUGAAGAUCUGGACAAUGUCUUUGUUACUAGACUGGG
CAUUAGUUCCUUAAUAAGAGAGGCAAUGAGUGCAUAUAAUAGUCAUGAA
GAGGGCAGAUUAGUUUACCGAUAUGGAGGAGAACCUGUUGGAUCCUUUG
UUCAGCCCUGUUUGAGGCCUUUAAUGCCAGCUAUUGCACAUGCCCUGUU
UAUGGAUAUUACGCAUGAUAAUGAGUGUCCUAUUGUGCAUAGAUCAGCG
UAUGAUGCUCUUCCAAGUACUACAAUUGUUUCUAUGGCAUGUUGUGCUA
GUGGAAGUACAAGAGGCUAUGAUGAAUUAGUGCCUCAUCAGAUUUCAGU
GGUUUCUGAAGAACGGUUUUACACUAAGUGGAAUCCUGAAGCAUUGCCU
UCAAACACAGGUGAAGUUAAUUUCCAAAGCGGCAUUAUUGCAGCCAGGU
GUGCUAUCAGUAAACUUCAUCAGGAGCUUGGAGCCAAGGGUUUUAUUCA
GGUGUAUGUGGAUCAAGUUGAUGAAGACAUAGUGGCAGUAACAAGACAC
UCACCUAGCAUCCAUCAGUCUGUUGUGGCUGUAUCUAGAACUGCUUUCA
GGAAUCCCAAGACUUCAUUUUACAGCAAGGAAGUGCCUCAAAUGUGCAU
CCCUGGCAAAAUUGAAGAAGUAGUUCUUGAAGCUAGAACUAUUGAGAGA
AACACGAAACCUUAUAGGAAGGAUGAGAAUUCAAUCAAUGGAACACCAG
AUAUCACAGUAGAAAUUAGAGAACAUAUUCAGCUUAAUGAAAGUAAAAU
UGUUAAACAAGCUGGAGUUGCCACAAAAGGGCCCAAUGAAUAUAUUCAA
GAAAUAGAAUUUGAAAACUUGUCUCCAGGAAGUGUUAUUAUAUUCAGAG
UUAGUCUUGAUCCACAUGCACAAGUCGCUGUUGGAAUUCUUCGAAAUCA
UCUGACACAAUUCAGUCCUCACUUUAAAUCUGGCAGCCUAGCUGUUGAC
AAUGCAGAUCCUAUAUUAAAAAUUCCUUUUGCUUCUCUUGCCUCCAGAU
UAACUUUGGCUGAGCUAAAUCAGAUCCUUUACCGAUGUGAAUCAGAAGA
AAAGGAAGAUGGUGGAGGGUGCUAUGACAUACCAAACUGGUCAGCCCUU
AAAUAUGCAGGUCUUCAAGGUUUAAUGUCUGUAUUGGCAGAAAUAAGAC
CAAAGAAUGACUUGGGGCAUCCUUUUUGUAAUAAUUUGAGAUCUGGAGA
UUGGAUGAUUGACUAUGUCAGUAACCGGCUUAUUUCACGAUCAGGAACU
AUUGCUGAAGUUGGUAAAUGGUUGCAGGCUAUGUUCUUCUACCUGAAGC
AGAUCCCACGUUACCUUAUCCCAUGUUACUUUGAUGCUAUAUUAAUUGG
UGCAUAUACCACUCUUCUGGAUACAGCAUGGAAGCAGAUGUCAAGCUUU
GUUCAGAAUGGUUCAACCUUUGUGAAACACCUUUCAUUGGGUUCAGUUC
AACUGUGUGGAGUAGGAAAAUUCCCUUCCCUGCCAAUUCUUUCACCUGC
CCUAAUGGAUGUACCUUAUAGGUUAAAUGAGAUCACAAAAGAAAAGGAG
CAAUGUUGUGUUUCUCUAGCUGCAGGCUUACCUCAUUUUUCUUCUGGUA
UUUUCCGCUGCUGGGGAAGGGAUACUUUUAUUGCACUUAGAGGUAUACU
GCUGAUUACUGGACGCUAUGUAGAAGCCAGGAAUAUUAUUUUAGCAUUU
GCGGGUACCCUGAGGCAUGGUCUCAUUCCUAAUCUACUGGGUGAAGGAA
UUUAUGCCAGAUACAAUUGUCGGGAUGCUGUGUGGUGGUGGCUGCAGUG
UAUCCAGGAUUACUGUAAAAUGGUUCCAAAUGGUCUAGACAUUCUCAAG
UGCCCAGUUUCCAGAAUGUAUCCUACAGAUGAUUCUGCUCCUUUGCCUG
CUGGCACACUGGAUCAGCCAUUGUUUGAAGUCAUACAGGAAGCAAUGCA
AAAACACAUGCAGGGCAUACAGUUCCGAGAAAGGAAUGCUGGUCCCCAG
AUAGAUCGAAACAUGAAGGACGAAGGUUUUAAUAUAACUGCAGGAGUUG
AUGAAGAAACAGGAUUUGUUUAUGGAGGAAAUCGUUUCAAUUGUGGCAC
AUGGAUGGAUAAAAUGGGAGAAAGUGACAGAGCUAGAAACAGAGGAAUC
CCAGCCACACCAAGAGAUGGGUCUGCUGUGGAAAUUGUGGGCCUGAGUA
AAUCUGCUGUUCGCUGGUUGCUGGAAUUAUCCAAAAAAAAUAUUUUCCC
UUAUCAUGAAGUCACAGUAAAAAGACAUGGAAAGGCUAUAAAGGUCUCA
UAUGAUGAGUGGAACAGAAAAAUACAAGACAACUUUGAAAAGCUAUUUC
AUGUUUCCGAAGACCCUUCAGAUUUAAAUGAAAAGCAUCCAAAUCUGGU
UCACAAACGUGGCAUAUACAAAGAUAGUUAUGGAGCUUCAAGUCCUUGG
UGUGACUAUCAGCUCAGGCCUAAUUUUACCAUAGCAAUGGUUGUGGCCC
CUGAGCUCUUUACUACAGAAAAAGCAUGGAAAGCUUUGGAGAUUGCAGA
AAAAAAAUUGCUUGGUCCCCUUGGCAUGAAAACUUUAGAUCCAGAUGAU
AUGGUUUACUGUGGAAUUUAUGACAAUGCAUUAGACAAUGACAACUACA
AUCUUGCUAAAGGUUUCAAUUAUCACCAAGGACCUGAGUGGCUGUGGCC
UAUUGGGUAUUUUCUUCGUGCAAAAUUAUAUUUUUCCAGAUUGAUGGGC
CCGGAGACUACUGCAAAGACUAUAGUUUUGGUUAAAAAUGUUCUUUCCC
GACAUUAUGUUCAUCUUGAGAGAUCCCCUUGGAAAGGACUUCCAGAACU
GACCAAUGAGAAUGCCCAGUACUGUCCUUUCAGCUGUGAAACACAAGCC
UGGUCAAUUGCUACUAUUCUUGAGACACUUUAUGAUUU{tilde over (A)}{tilde over
(U)}{tilde over (A)}G (SEQ ID NO: 151) A GGGACACAG AAACAGA CGAA AC
C GAACGAAA GGAGAA AC GGAAAAGACCC C CAGAC GAACAAGGG A GAGC ACAG CCGA
AGGCCCAAC ACAGGGAAAAGCAG ACCG G A ACAA A ACCCA CC GGAGAAACA AA
AGAGAAAAA CCG C C GGA GGGAAAA CCAACAGAAAGAGAAGA GA C GA AAA A C G
AAAC AA C GCAACAA C GG CA CAG A A C C CAAGGAAA GAGAAAAG GG GGAGG
ACA AG G GGACCCCA ACG G GG GC GA AA CA G GC ACCC GGAC G G AC C
CAGACA AGC AAG G GGGACC GA GAA GGGAAAGCAGAC AGGG GCAAAAGAA CAGGC
ACAACA GA CA ACCCCA GCAGAC C GGAC A C AGG CA GC AC CCC GCCAA CAG
AGAA AAA CC GAC CAAGACC AA AGAAAG A ACC GGAA GA G GGACAGC AG
GGAAAAA AAA AAAGGAA GGAA G A G A AC GA G G C ACAA CA AC GC GC AA AG
AAA GGA CCAGGAACA CCAGAA G GCC A AA CU G GAANC CCACAC AAAACC GCC
GGG C AGACAGAGCA C GGCG CC G GA G GCAGAAGGGAAA ACAAAGAAAA GGGAA ACC
GC GA GAAAA GA CACCA A GAA CCA CC GAAAAA AA GGGAGGA A CCAAAGC AAAC
C GGGA A CCAAG AGA G CAACAAAGCGG GAGCA AGAAGAC C ACACAAGAAAA
AGGCGAG AACCAAG C GA CCAAACCAA CACC ACGA A CAAGA CC GAA ACAGACGG
GGC G AC G AGA A GAACA GCAC AACGAC CA ACCACA GACAAGGGG CCAGCAGCAA
GAAGAA GC G AA GG CA AAAAGAA GGA GGAA AAA CAGAGAAGCA CGAC CA AAC A
CA CAGGAACA GGCAG AA GCC GGGAAA G G A GAACGAC GGC G GCCA GG CCAAAAC
AGGACC G CAC AGAAAGCA CC AG A CCAGG A AC CCCA GAAGAGA AGAC C CCA
GGAA GAA C A GA CA C GCCAAA AAAGC G C GA GGCACACA
A GGA GGG AA GGGAGA GA CC C CGAAAC GC GAACCGG G CAGAAG ACC
AAGGAGAGAAC A GC GGGGAGACAG G AAA ACGC A GGGAA AAACCAGAGGAC G CC A
C C GG GCACACA GAAAAAA ACAC GAAA AAC GCAAC A CCAGGGA G ACG C GA AAC
GCCAC CAACACC C CACG AGC GAG AC A G GGA GC GC AGGAA GCAACCCAA A A G
AG AGC GAAC G CACAGGAAG GAAGA C GGACAA G C G AC A GAC GGGCA AG CC
AA AAGAGAGGCAA GAG GCA A AA AG CA GAAGAGGGCAGA AG ACCGA A
GGAGGAGAACC G GGA CC G CAGCCC G GAGGCC AA GCCAGC A G CACA GCCC G A
GGA A ACGCA GA AA GAG G CC A G GCA AGA CAGCG A GA GC C CCAAG AC
ACAA G C A GGCA G G GC AG GGAAG ACAAGAGGC A GA GAA AG GCC CA CAGA
CAG GG C GAAGAACGG ACAC AAG GGAA CC GAAGCA GCC CAAACACAGG GAAG AA
CCAAA GCGGCA A GCAGCCAGG G GC A CAG AAAC CA CAGGAGC GGAGCCAAGGG A
CAGG G A G GGA CAAG GA G AAGACA AG GGCAG AACAAGACAC CACC AGCA CCA
CAG C G G GGC G A C AGAAC GC CAGGAA CCCAAGAC CA ACAGCAAGGAAG GCC
CAAA G GCA CCC GGCAAAA GAAGAAG AG C GAAGC AGAAC A GAGAGAAACACGAAAC
C A AGGAAGGA GAGAA CAA CAA GGAACACCAGA A CAC AG AGAAA AGAGAACA A
CAGC AA GAAAG AAAA G A AACAAGC GGAG GCCACAAAAGGGCCCAA GAA A A
CAAGAA A AGAA GAAAAC G C CCAGGAAG G A A A CAGAG AG C GA CCACA
GCACAAG CGC G GGAA C CGAAA CA C GACACAA CAG CC CAC AAA C GGCAGCC
AGC G GACAA GCAGA CC A A AAAAA CC GC C C GCC CC AGA AAC GGC GAGC
AAA CAGA CC ACCGA G GAA C AGAAGAAAAGGAAGA GG GGAGGG GC A GACA
ACCAAAC GG CAGCCC AAA A GCAGG C CAAGG AA G C G A GGC AGAAA
AAGACCAAAGAA GAC GGGGCA CC G AA AA GAGA C GGAGA GGA GA GAC A G CAG
AACCGGC A CACGA CAGGAAC A GC GAAG GG AAA GG GCAG GC A G C C ACC
GAAGCAGA CCCACG ACC A CCCA G AC GA GC A A AA GG GCA A ACCAC C C GGA
A CAGCA GGAAGCAGA G CAAGC G CAGAA GG CAACC G GAAACACC CA GGG CAG
CAAC G G GGAG AGGAAAA CCC CCC GCCAA C CACC GCCC AA GGA G ACC A AGG
AAA GAGA CACAAAAGAAAAGGAGCAA G G G C AG C GCAGGC ACC CA C C GG A
CCGC GC GGGGA AGGGA AC A GCAC AGAGG A AC GC GA AC GGACGC A G
AGAAGCCAGGAA A A AGCA GCGGG ACCC GAGG CA GG C CA CC AA C AC GGG
GAAGGA A GCCAGA A CAA G CGGGA GC G G GG GG GGC GCAG G A CCAGGA AC G
AAAA GG CCAAA GG C AGACA C CAAG GCCCA G CCAGAA G A CC ACAGA GA C GC
CC GCC GC G GCACAC GGA CAGCCA G GAAG CA ACAGGAAGCAA GCAA AAACACA
GCAGGGCA ACAG CCGAGAAAGGAA GC GG CCCCAG A AGA CGAAACA GAAGGACGAAGG
AA A AAC GCAGGAG GA GAAGAAACAGGA G A GGAGGAAA CG CAA G GGC ACA GGA
GGA AAAA GGGAGAAAG GACAGAGC AGAAACAGAGGAA CCCAGCCACACCAAGAGA GGG C
GC G GGAAA G GGGCC GA G AAA C GC G CGC GG GC GGAA A CCAAAAAAAA CCC
A CA GAAG CACAG AAAAAGACA GGAAAGGC A AAA GG C CA A GA GAG
GGAACAGAAAAA ACAAGACAAC GAAAA GC A CA G CCGAAGACCC CAGA AAA
GAAAAGCA CC AAA C GG CACAAACG GGCA A ACAAAGA AG A GGAGC CAAG CC GG
G GAC A CAGC CAGGCC AA ACCA AGCAA GG G GGCCCC GAGC C AC
ACAGAAAAAGCA GGAAAGC GGAGA GCAGAAAAAAAA GC GG CCCC GGCA GAAA AC AGA
CCAGA GA A GG AC G GGA A GACAA G CA AGACAA GACAAC ACAA C GC AAAGG
CAA A CACC AAGGACC GAG GGC G GGCC A GGG A CG GCAAAA A A CCAGA GA
GGGCCCGGAGAC AC GCAAAGAC A AG GG AAAAA G C CCCGACA A G CA C GAGAGA
C CCC GGAAAGGAC CCAGAAC GACCAA GAGAA GCCCAG AC G CC CAGC G
GAAACACAAGCC GG CAA GC AC A C GAG ACAC A GA A AG
Example 22
mUNA Oligomer Expressing Human Protein S (alpha) (PROS1)
[0492] In this example, the structures of mUNA molecules for use in
expressing human protein S (alpha) (PROS1) are shown.
[0493] Human protein S (alpha) is associated with Protein S
deficiency, thrombosis, and arterial occlusive disease.
[0494] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human protein S (alpha). The complete
mUNA molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR upstream
of the sequence below, and a 3' UTR and polyA tail (SEQ ID Nos: 4
to 12) downstream of the sequence below, each of which corresponds
to the structure of the native mRNA of human protein S (alpha).
TABLE-US-00017 Human protein S (alpha) is accession NM_001314077.1.
(SEQ ID NO: 152) A{tilde over (U)}{circumflex over (G)}{tilde over
(A)}GGGUCCUGGGUGGGCGCUGCGGGGCGCUGCUGGCGUGUCUCCUC
CUAGUGCUUCCCGUCUCAGAGGCAAACUUUUGUUUAUAUUUUAGAAAUG
AUUUUAUAUACAACCGUGCAUGCAUUUCUGUAUUGGUCGGCUUAUCUGG
AUGCAAUCUUUUUUCUAUUCUAUAUGCUUUUUGAAAGCAACAGGCUUCA
CAAGUCCUGGUUAGGAAGCGUCGUGCAAAUUCUUUACUUGAAGAAACCA
AACAGGGUAAUCUUGAAAGAGAAUGCAUCGAAGAACUGUGCAAUAAAGA
AGAAGCCAGGGAGGUCUUUGAAAAUGACCCGGAAACGGAUUAUUUUUAU
CCAAAAUACUUAGUUUGUCUUCGCUCUUUUCAAACUGGGUUAUUCACUG
CUGCACGUCAGUCAACUAAUGCUUAUCCUGACCUAAGAAGCUGUGUCAA
UGCCAUUCCAGACCAGUGUAGUCCUCUGCCAUGCAAUGAAGAUGGAUAU
AUGAGCUGCAAAGAUGGAAAAGCUUCUUUUACUUGCACUUGUAAACCAG
GUUGGCAAGGAGAAAAGUGUGAAUUUGACAUAAAUGAAUGCAAAGAUCC
CUCAAAUAUAAAUGGAGGUUGCAGUCAAAUUUGUGAUAAUACACCUGGA
AGUUACCACUGUUCCUGUAAAAAUGGUUUUGUUAUGCUUUCAAAUAAGA
AAGAUUGUAAAGAUGUGGAUGAAUGCUCUUUGAAGCCAAGCAUUUGUGG
CACAGCUGUGUGCAAGAACAUCCCAGGAGAUUUUGAAUGUGAAUGCCCC
GAAGGCUACAGAUAUAAUCUCAAAUCAAAGUCUUGUGAAGAUAUAGAUG
AAUGCUCUGAGAACAUGUGUGCUCAGCUUUGUGUCAAUUACCCUGGAGG
UUACACUUGCUAUUGUGAUGGGAAGAAAGGAUUCAAACUUGCCCAAGAU
CAGAAGAGUUGUGAGGUUGUUUCAGUGUGCCUUCCCUUGAACCUUGACA
CAAAGUAUGAAUUACUUUACUUGGCGGAGCAGUUUGCAGGGGUUGUUUU
AUAUUUAAAAUUUCGUUUGCCAGAAAUCAGCAGAUUUUCAGCAGAAUUU
GAUUUCCGGACAUAUGAUUCAGAAGGCGUGAUACUGUACGCAGAAUCUA
UCGAUCACUCAGCGUGGCUCCUGAUUGCACUUCGUGGUGGAAAGAUUGA
AGUUCAGCUUAAGAAUGAACAUACAUCCAAAAUCACAACUGGAGGUGAU
GUUAUUAAUAAUGGUCUAUGGAAUAUGGUGUCUGUGGAAGAAUUAGAAC
AUAGUAUUAGCAUUAAAAUAGCUAAAGAAGCUGUGAUGGAUAUAAAUAA
ACCUGGACCCCUUUUUAAGCCGGAAAAUGGAUUGCUGGAAACCAAAGUA
UACUUUGCAGGAUUCCCUCGGAAAGUGGAAAGUGAACUCAUUAAACCGA
UUAACCCUCGUCUAGAUGGAUGUAUACGAAGCUGGAAUUUGAUGAAGCA
AGGAGCUUCUGGAAUAAAGGAAAUUAUUCAAGAAAAACAAAAUAAGCAU
UGCCUGGUUACUGUGGAGAAGGGCUCCUACUAUCCUGGUUCUGGAAUUG
CUCAAUUUCACAUAGAUUAUAAUAAUGUAUCCAGUGCUGAGGGUUGGCA
UGUAAAUGUGACCUUGAAUAUUCGUCCAUCCACGGGCACUGGUGUUAUG
CUUGCCUUGGUUUCUGGUAACAACACAGUGCCCUUUGCUGUGUCCUUGG
UGGACUCCACCUCUGAAAAAUCACAGGAUAUUCUGUUAUCUGUUGAAAA
UACUGUAAUAUAUCGGAUACAGGCCCUAAGUCUAUGUUCCGAUCAACAA
UCUCAUCUGGAAUUUAGAGUCAACAGAAACAAUCUGGAGUUGUCGACAC
CACUUAAAAUAGAAACCAUCUCCCAUGAAGACCUUCAAAGACAACUUGC
CGUCUUGGACAAAGCAAUGAAAGCAAAAGUGGCCACAUACCUGGGUGGC
CUUCCAGAUGUUCCAUUCAGUGCCACACCAGUGAAUGCCUUUUAUAAUG
GCUGCAUGGAAGUGAAUAUUAAUGGUGUACAGUUGGAUCUGGAUGAAGC
CAUUUCUAAACAUAAUGAUAUUAGAGCUCACUC
AUGUCCAUCAGUUUGGAAAAAGACAAAGAAUUCU{tilde over (U)}{tilde over
(U)}{tilde over (A)}A (SEQ ID NO: 153) A GAGGG CC GGG GGGCGC
GCGGGGCGC GC GGCG G C CC CC AG GC CCCG C CAGAGGCAAAC G A A AGAAA GA
A A ACAACCG GCA GCA C G A GG CGGC A C GGA GCAA C A C A A GC G
CAAAGCAACA GGC CACAAG CC GG AGGAAGCG CG GCAAA C AC G
AAGAAACCAAACAGGG AA C GAAAGAGAA GCA CGAAGAAC G G CAA
AAAGAAGAAGCCAGGGAGG C GAAAA GACCCGGAAACGGA A A CCAAAA AC AG G C CGC
C CAAAC GGG A CAC GC GCACG CAG CAAC AA GC A CC GACC A AGAAGC G G
CAA GCCA CCAGACCAG G AG CC C GCCA GC AA GAAGA GGA A A GAGC GCAAAGA
GGAAAAGC C AC GCAC G AAACCAGG GGCAAGGAGAAAAG G GAA GACA AAA GAA
GCAAAGA CCC CAAA A AAA GGAGG GCAG CAAA G GA AA ACACC GGAAG ACCAC G
CC G AAAAA GG G A GC CAAA AAGAAAGA G AAAGA G GGA GAA GC C
GAAGCCAAGCA G GGCACAGC G G GCAAGAACA CCCAGGAGA GAA G GAA
GCCCCGAAGGC ACAGA A AA C CAAA CAAAG C G GAAGA A AGA GAA GC C
GAGAACA G G GC CAGC NG G CAA ACCC GGAGG ACAC GC A G GA GGGAAGAAA
GGA CAAAC GCCCAAGA CAGAAGAG G GAGG G CAG G GCC CCC GAACC GACACAAAG
A GAA AC AC GGC GGAGCAG GCAGGGG G A AAAA CG GCCAGA AA CAGCAGA
CAGCAGAA GA CCGGACA A GA CAGA AGGCG GA AC G ACGCAGAA C A CGA CAC
CAGCG GGC C C GA GCAC CG GG GGAAAGA GAAG CAGC AAGAA GAA CA ACA
CCAAAA CACAAC GGAGG GA G A AA AA GG C A GGAA A GG G C G GGAAGAA
AGAACA AG A AGCA AAAA AGC AAAGAAGC G GA GGA A AAA AAACC GGACCCC
AAGCCGGAAAA GGA GC GGAAACCAAAG A AC GCAGG A CCC CGGAAAG GGAAAG GAAC
CA AAACCGA AACCC C G C AGA GGA G A ACGAAGC GGAA GA GAAGCAAGGAGC
GGAA AAAGGAAA A CAAGAAAAACAAAA AAGCA GCC G G AC G GGAGAAGGGC CC AC
A CC GG C GGAA GC CAA CACA AGA A AA AA G A CCAG GC GAGGG GGCA G AAA
G GACC GAA A CG CCA CCACGGGCAC GG G A G C GCC GG C GG AACAACACAG
GCCC GC G G CC GG GGAC CCACC C GAAAAA CACAGGA A C G A C G GAA AA AC
G AA A A CGGA ACAGGCCC AAG C A G CCGA CAAC AA C CA C GGAA AGAG
CAACAGAAACAA C GGAG G CGA CACCAC AAAA AGAAACCA C CCCA GAAGACC
CAAAGACAAC GCCG C GGACAAAGCAA GAAAGCAAAAG GGCCACA ACC GGG GGCC
CCAGA G CCA CAG GCCACACCAG GAA GCC A AA GGC GCA GGAAG GAA A AA GG G
ACAG GGA C GGA GAAGCCA C AAACA AA GA A AGAGC CAC CA G CCA CA G
GGAAAAAGACAAAGAA C AA
Example 23
mUNA Oligomer Expressing Human Pyruvate Kinase, Liver and RBC
(PKLR)
[0495] In this example, the structures of mUNA molecules for use in
expressing human pyruvate kinase, liver and RBC (PKLR) are
shown.
[0496] Human pyruvate kinase, liver and RBC (PKLR) is associated
with chronic hereditary nonspherocytic hemolytic anemia.
[0497] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human pyruvate kinase, liver and RBC
(PKLR). The complete mUNA molecule comprises a 5' cap (m7GpppGm),
and a 5'-UTR upstream of the sequence below, and a 3' UTR and polyA
tail (SEQ ID Nos: 4 to 12) downstream of the sequence below, each
of which corresponds to the structure of the native mRNA of human
pyruvate kinase, liver and RBC (PKLR).
TABLE-US-00018 Human pyruvate kinase, liver and RBC (PKLR) is
accession NM_000298.5. (SEQ ID NO: 154) A{tilde over
(U)}{circumflex over (G)}{tilde over
(U)}CGAUCCAGGAGAACAUAUCAUCCCUGCAGCUUCGGUCAUGGGUC
UCUAAGUCCCAAAGAGACUUAGCAAAGUCCAUCCUGAUUGGGGCUCCA
GGAGGGCCAGCGGGGUAUCUGCGGCGGGCCAGUGUGGCCCAACUGACC
CAGGAGCUGGGCACUGCCUUCUUCCAGCAGCAGCAGCUGCCAGCUGCU
AUGGCAGACACCUUCCUGGAACACCUCUGCCUACUGGACAUUGACUCC
GAGCCCGUGGCUGCUCGCAGUACCAGCAUCAUUGCCACCAUCGGGCCA
GCAUCUCGCUCCGUGGAGCGCCUCAAGGAGAUGAUCAAGGCCGGGAUG
AACAUUGCGCGACUCAACUUCUCCCACGGCUCCCACGAGUACCAUGCU
GAGUCCAUCGCCAACGUCCGGGAGGCGGUGGAGAGCUUUGCAGGUUCC
CCACUCAGCUACCGGCCCGUGGCCAUCGCCCUGGACACCAAGGGACCG
GAGAUCCGCACUGGGAUCCUGCAGGGGGGUCCAGAGUCGGAAGUGGAG
CUGGUGAAGGGCUCCCAGGUGCUGGUGACUGUGGACCCCGCGUUCCGG
ACGCGGGGGAACGCGAACACCGUGUGGGUGGACUACCCCAAUAUUGUC
CGGGUCGUGCCGGUGGGGGGCCGCAUCUACAUUGACGACGGGCUCAUC
UCCCUAGUGGUCCAGAAAAUCGGCCCAGAGGGACUGGUGACCCAAGUG
GAGAACGGCGGCGUCCUGGGCAGCCGGAAGGGCGUGAACUUGCCAGGG
GCCCAGGUGGACUUGCCCGGGCUGUCCGAGCAGGACGUCCGAGACCUG
CGCUUCGGGGUGGAGCAUGGGGUGGACAUCGUCUUUGCCUCCUUUGUG
CGGAAAGCCAGCGACGUGGCUGCCGUCAGGGCUGCUCUGGGUCCGGAA
GGACACGGCAUCAAGAUCAUCAGCAAAAUUGAGAACCACGAAGGCGUG
AAGAGGUUUGAUGAAAUCCUGGAGGUGAGCGACGGCAUCAUGGUGGCA
CGGGGGGACCUAGGCAUCGAGAUCCCAGCAGAGAAGGUUUUCCUGGCU
CAGAAGAUGAUGAUUGGGCGCUGCAACUUGGCGGGCAAGCCUGUUGUC
UGUGCCACACAGAUGCUGGAGAGCAUGAUUACCAAGCCCCGGCCAACG
AGGGCAGAGACAAGCGAUGUCGCCAAUGCUGUGCUGGAUGGGGCUGAC
UGCAUCAUGCUGUCAGGGGAGACUGCCAAGGGCAACUUCCCUGUGGAA
GCGGUGAAGAUGCAGCAUGCGAUUGCCCGGGAGGCAGAGGCCGCAGUG
UACCACCGGCAGCUGUUUGAGGAGCUACGUCGGGCAGCGCCACUAAGC
CGUGAUCCCACUGAGGUCACCGCCAUUGGUGCUGUGGAGGCUGCCUUC
AAGUGCUGUGCUGCUGCCAUCAUUGUGCUGACCACAACUGGCCGCUCA
GCCCAGCUUCUGUCUCGGUACCGACCUCGGGCAGCAGUCAUUGCUGUC
ACCCGCUCUGCCCAGGCUGCCCGCCAGGUCCACUUAUGCCGAGGAGUC
UUCCCCUUGCUUUACCGUGAACCUCCAGAAGCCAUCUGGGCAGAUGAU
GUAGAUCGCCGGGUGCAAUUUGGCAUUGAAAGUGGAAAGCUCCGUGGC
UUCCUCCGUGUUGGAGACCUGGUGAUUGUGGUGACAGGCUGGCGACCU
GGCUCCGGCUACACCAACAUCAUGCGGGUGCUAAGCAUAUC{circumflex over
(C)}{tilde over (U)}{circumflex over (G)}A (SEQ ID NO: 155) A G CGA
CCAGGAGAACA A CA CCC GCAGC CGG CA GGG C C AAG CCCA AAGAGAC AGCAAAG
CCA CC GA GGGGC CCAGGAGGGCCAGCGGGG A C GC GGCGGGCCAG G GGCCCAAC
GACCCAGGAGC GGGCAC GCC C CCAGCAGCAG CAGC GCCAGC GC A GGCAGACACC CC
GGAACACC C GCC AC GGACA GA C CCGAGCCCG GGC GC CGCAG ACCAGCA CA
GCCACCA CGGGCCAGCA C C GC CCG GGAGCGCC CAAGGAGA GA CAAGGCCGGGA
GAACA GCGCGAC CAAC C CCCACGGC CCCACGAG ACCA GC GAG CCA CGCCAACG
CCGGGAGGCGG GGAGAGC GCAGG CCCCAC CAGC ACCGGCCCG GGCCA CGCCC
GGACACCA AGGGACCGGAGA CCGCAC GGGA CC GCAGGGGGG CCAGAG CGGAAG GGAGC
G G GAAGGGC CCCAGG GC GG GAC G GGACCCCGCG CCGGACGCGGGGGAACGC
GAACACCG G GGG GGAC ACCCCAA A G CCGGG CG GCCGG GGGGGGCCGCA C ACA
GACGACGGGC CA C CCC AG GG CCAGAAAA CGGCCCAGAGGGAC G G GACCCAAG
GGAGAACGGCGGCG CC GGGCAGCCGGAAGGGCG GAAC GCCAGG GGCCCAGG GGAC
GCCCGGGC G CCGAGCAGGACG CCGAGACC GCGC CGGGG GGAGCA GGGG GGACA CG C
GCC CC G GCGGAAAGCCAGCGACG GGC GCCG CAGGGC GC C GGG
CCGGAAGGACACGGCA CAAGA CA CAGCAAAA GA GAACCACGAAGGCG GAAGAGG GA
GAAA CC GGAGG GAGCGACGGCA CA GG GGCACGGGGGGACC AGGCA CGAGA
CCCAGCAGAGAAGG CC GGC CAGAAG A GA GA GGGCGC GCAAC GGCGGGCAAGCC G G
C G GCCACACAGA GC GGAGAGCA GA ACCAAGCCCCGGCCAACGAGGGCAGAGACAAGCGA G
CGCCAA G C G GC GGA GGGGC GAC GCA CA GC G CAGGGGAGAC GCCAAGGGCAAC C
CC G GGAAGCGG GAAGA GCAGCA GCGA GCCCGGGAGGCAGAGGCCGCAG G A
CCACCGGCAGC G GAGGAGC ACG CGGGCAGCGCCAC AAGCCG GA CCCAC G AGG
CACCGCCA GG GC G GGAGGC GCC CAAG GC G GC GC GCCA CA G GC GACCACAAC
GGCCGC CAGCCCAGC C G C CGG ACCGACC CGGGCAGC AG CA GC G CACCCGC C
GCCCAGGC GCCCGCCAGG CCAC A GCCGAGGAG C CCCC GC ACCG GAACC
CCAGAAGCCA C GGGCAGA GA G AGA CGC CGGG GCAA GGCA GAAAG GGAAAGC CCG
GGC CC CCG G GGAGACC GG GA G GG GACAGGC GGCGACC GGC CCGGC ACACCAACA
CA GCGGG GC AAGCA A CC GA
Example 24
mUNA Oligomer Expressing Human Phenylalanine Hydroxylase
[0498] In this example, the structures of mUNA molecules for use in
expressing human phenylalanine hydroxylase are shown.
[0499] Human phenylalanine hydroxylase is associated with
phenylketonuria.
[0500] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the open reading
frame of the native mRNA of human phenylalanine hydroxylase. The
complete mUNA molecule comprises a 5' cap (m7GpppGm), and a 5'-UTR
upstream of the sequence below, and a 3' UTR and polyA tail (SEQ ID
NOs: 4 to 12) downstream of the sequence below, each of which
corresponds to the structure of the native mRNA of human
phenylalanine hydroxylase.
TABLE-US-00019 Human phenylalanine hydroxylase is accession
NM_000277.1. (SEQ ID NO: 156) A{tilde over (U)}{circumflex over
(G)}{tilde over
(U)}CCACUGCGGUCCUGGAAAACCCAGGCUUGGGCAGGAAACUCUCUGACUUUGGACA
GGAAACAAGCUAUAUUGAAGACAACUGCAAUCAAAAUGGUGCCAUAUCACUGAUCUUCU
CACUCAAAGAAGAAGUUGGUGCAUUGGCCAAAGUAUUGCGCUUAUUUGAGGAGAAUGAU
GUAAACCUGACCCACAUUGAAUCUAGACCUUCUCGUUUAAAGAAAGAUGAGUAUGAAUU
UUUCACCCAUUUGGAUAAACGUAGCCUGCCUGCUCUGACAAACAUCAUCAAGAUCUUGA
GGCAUGACAUUGGUGCCACUGUCCAUGAGCUUUCACGAGAUAAGAAGAAAGACACAGUG
CCCUGGUUCCCAAGAACCAUUCAAGAGCUGGACAGAUUUGCCAAUCAGAUUCUCAGCUA
UGGAGCGGAACUGGAUGCUGACCACCCUGGUUUUAAAGAUCCUGUGUACCGUGCAAGAC
GGAAGCAGUUUGCUGACAUUGCCUACAACUACCGCCAUGGGCAGCCCAUCCCUCGAGUG
GAAUACAUGGAGGAAGAAAAGAAAACAUGGGGCACAGUGUUCAAGACUCUGAAGUCCUU
GUAUAAAACCCAUGCUUGCUAUGAGUACAAUCACAUUUUUCCACUUCUUGAAAAGUACU
GUGGCUUCCAUGAAGAUAACAUUCCCCAGCUGGAAGACGUUUCUCAAUUCCUGCAGACU
UGCACUGGUUUCCGCCUCCGACCUGUGGCUGGCCUGCUUUCCUCUCGGGAUUUCUUGGG
UGGCCUGGCCUUCCGAGUCUUCCACUGCACACAGUACAUCAGACAUGGAUCCAAGCCCA
UGUAUACCCCCGAACCUGACAUCUGCCAUGAGCUGUUGGGACAUGUGCCCUUGUUUUCA
GAUCGCAGCUUUGCCCAGUUUUCCCAGGAAAUUGGCCUUGCCUCUCUGGGUGCACCUGA
UGAAUACAUUGAAAAGCUCGCCACAAUUUACUGGUUUACUGUGGAGUUUGGGCUCUGCA
AACAAGGAGACUCCAUAAAGGCAUAUGGUGCUGGGCUCCUGUCAUCCUUUGGUGAAUUA
CAGUACUGCUUAUCAGAGAAGCCAAAGCUUCUCCCCCUGGAGCUGGAGAAGACAGCCAU
CCAAAAUUACACUGUCACGGAGUUCCAGCCCCUGUAUUACGUGGCAGAGAGUUUUAAUG
AUGCCAAGGAGAAAGUAAGGAACUUUGCUGCCACAAUACCUCGGCCCUUCUCAGUUCGC
UACGACCCAUACACCCAAAGGAUUGAGGUCUUGGACAAUACCCAGCAGCUUAAGAUUUU
GGCUGAUUCCAUUAACAGUGAAAUUGGAAUCCUUUGCAGUGCCCUCCAGAAAAUAAA{circumflex
over (G)}{tilde over (U)} {tilde over (A)}A (SEQ ID NO: 157)
AU{circumflex over (G)}UCCACUGCGGUCCUGGA{tilde over
(A)}AACCCAGGCUUGGGCAG{circumflex over (G)}AAACUCUCUGACUUUGG{tilde
over (A)}CA GGAAACAAGCUAUAUUGAAGACAACUGCAAUCA{tilde over
(A)}AAUGGUGCCAUAUCACU{circumflex over (G)}AUCUUCU CACUCAAAGA{tilde
over (A)}GAAGUUGGUGCAUUGGC{circumflex over
(C)}AAAGUAUUGCGCUUAUU{tilde over (U)}GAGGAGAAUGAU GUAAA{circumflex
over (C)}CUGACCCACAUUGAAUC{tilde over
(U)}AGACCUUCUCGUUUAAA{circumflex over (G)}AAAGAUGAGUAUGAAUU {tilde
over (U)}UUCACCCAUUUGGAUAA{tilde over (A)}CGUAGCCUGCCU{circumflex
over (G)}CUCUGACAAACAUCAUCAAGAU{circumflex over (C)}UUGA
GGCAUGACAUUGG{tilde over (U)}GCCACUGUCCAUGAGCU{tilde over
(U)}UCACGAGAUAAGAAGAA{tilde over (A)}GACACAGUG CCCUGGUU{circumflex
over (C)}CCAAGAACCAUUCAAGA{circumflex over
(G)}CUGGACAGAUUUGCCAA{tilde over (U)}CAGAUUCUCAGCUA UGG{tilde over
(A)}GCGGAACUGGAUGCUGA{circumflex over (C)}CACCCUGGUUUUAAAGA{tilde
over (U)}CCUGUGUACCGUGCAAG{tilde over (A)}C
GGAAGCAGUUUGCUGA{circumflex over (C)}AUUGCCUACAACUACCG{circumflex
over (C)}CAUGGGCAGCCCAUCCC{tilde over (U)}CGAGUG
GAAUACAUGGAGGAAGAAAAGAAAACAUGGGGCACAGUGUUCAAGAC{tilde over
(U)}CUGAAGUCCUU GUAUAA{tilde over (A)}ACCCAUGCUUGCUAUGA{circumflex
over (G)}UACAAUCACAUUUUUCC{tilde over (A)}CUUCUUGAAAAGUACU G{tilde
over (U)}GGCUUCCAUGAAGAUAA{circumflex over
(C)}AUUCCCCAGCUGGAAGA{circumflex over
(C)}GUUUCUCAAUUCCUGCA{circumflex over (G)}ACU
UGCACUGGUUUCCG{circumflex over (C)}CUCCGACCUGUGGCUGG{circumflex
over (C)}CUGCUUUCCUCUCGGGA{tilde over (U)}UUCUUGGG
UGGCCUGGC{circumflex over (C)}UUCCGAGUCUUCCACUG{circumflex over
(C)}ACACAGUACAUCAGACA{tilde over (U)}GGAUCCAAGCCCA UGUA{tilde over
(U)}ACCCCCGAACCUGACAU{circumflex over (C)}UGCCAUGAGCUGUUGGG{tilde
over (A)}CAUGUGCCCUUGUUUUC{tilde over (A)}
GAUCGCAGCUUUGCCCA{circumflex over (G)}UUUUCCCAGGAAAUUGG{circumflex
over (C)}CUUGCCUCUCUGGGUGC{tilde over (A)}CCUGA UGAAUACAUUGA{tilde
over (A)}AAGCUCGCCACAAUUUA{circumflex over
(C)}UGGUUUACUGUGGAGUU{tilde over (U)}GGGCUCUGCA AACAAGG{tilde over
(A)}GACUCCAUAAAGGCAUA{tilde over (U)}GGUGCUGGGCUCCUGUC{tilde over
(A)}UCCUUUGGUGAAUUA CA{circumflex over
(G)}UACUGCUUAUCAGAGAA{circumflex over
(G)}CCAAAGCUUCUCCCCCU{circumflex over
(G)}GAGCUGGAGAAGACAGC{circumflex over (C)}AU
CCAAAAUUACACUGU{circumflex over (C)}ACGGAGUUCCAGCCCCU{circumflex
over (G)}UAUUACGUGGCAGAGAG{tilde over (U)}UUUAAUG
AUGCCAAGGA{circumflex over (G)}GAAAGUAAGGAACUUUG{circumflex over
(C)}UGCCACAAUACCUCGGC{circumflex over (C)}CUUCUCAGUUCG CUACG{tilde
over (A)}CCCAUACACCCAAAGGA{tilde over
(U)}UGAGGUCUUGGACAAUA{circumflex over (C)}CCAGCAGCUUAAGAUUU {tilde
over (U)}GGCUGAUUCCAUUAACA{circumflex over
(G)}UGAAAUUGGAAUCCUUU{circumflex over (G)}CAGUGCCCUCCAGAAAA{tilde
over (U)}AAAG UAA (SEQ ID NO: 158) A G CCAC GCGG CC GGAAAACCCAGGC
GGGCAGGAAAC C C GAC GGACA GGAAACAAGC A A GAAGACAAC GCAA CAAAA GG
GCCA A CAC GA C C CAC CAAAGAAGAAG GG GCA GGCCAAAG A GCGC A GAGGAGAA
GA G AAACC GACCCACA GAA C AGACC C CG AAAGAAAGA GAG A GAA CACCCA GGA
AAACG AGCC GCC GC C GACAAACA CA CAAGA C GA GGCA GACA GG GCCAC G CCA
GAGC CACGAGA AAGAAGAAAGACACAG G CCC GG CCCAAGAACCA CAAGAGC GGACAGA
GCCAA CAGA C CAGC A GGAGCGGAAC GGA GC GACCACCC GG AAAGA CC G G ACCG
GCAAGAC GGAAGCAG GC GACA GCC ACAAC ACCGCCA GGGCAGCCCA CCC CGAG G
GAA ACA GGAGGAAGAAAAGAAAACA GGGGCACAG G CAAGAC C GAAG CC G A
AAAACCCA GC GC A GAG ACAA CACA CCAC C GAAAAG AC G GGC CCA GAAGA
AACA CCCCAGC GGAAGACG C CAA CC GCAGAC GCAC GG CCGCC CCGACC G GGC
GGCC GC CC C CGGGA C GGG GGCC GGCC CCGAG C CCAC GCACACAG ACA CAGACA
GGA CCAAGCCCA G A ACCCCCGAACC GACA C GCCA GAGC G GGGACA G GCCC G CA
GA CGCAGC GCCCAG CCCAGGAAA GGCC GCC C C GGG GCACC GA GAA ACA
GAAAAGC CGCCACAA AC GG AC G GGAG GGGC C GCA AACAAGGAGAC CCA AAAGGCA
A GG GC GGGC CC G CA CC GG GAA A CAG AC GC A CAGAGAAGCCAAAGC C
CCCCC GGAGC GGAGAAGACAGCCA CCAAAA ACAC G CACGGAG CCAGCCCC G A ACG
GGCAGAGAG AA G A GCCAAGGAGAAAG AAGGAAC GC GCCACAA ACC CGGCCC C CAG
CGC ACGACCCA ACACCCAAAGGA GAGG C GGACAA ACCCAGCAGC AAGA GGC GA CCA
AACAG GAAA GGAA CC GCAG GCCC CCAGAAAA AAAG AA
Example 25
mUNA Oligomer Translation Enhancer Based on TEV 5'UTR
[0501] In this example, the structures of mUNA molecules for
enhancing translational efficiency are shown.
[0502] The 5'-UTR of tobacco etch virus (TEV) is as follows:
TABLE-US-00020 (SEQ ID NO: 159)
UCAACACAACAUAUACAAAAACAAACGAAUCUCAAGCAAUCAAGCAUU
CUACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAA
UUUUCUGAAAAUUUUCACCAUUUACGAACGAUAGCC
[0503] The base sequences shown below are the portion of the mUNA
molecule that may correspond in functionality to the 5'-UTR of
tobacco etch virus (TEV). The complete mUNA molecule comprises a 5'
cap upstream of the sequence below (m7GpppGm), and a coding region
(CDS) of a protein of interest, a 3'-UTR, and a polyA tail (SEQ ID
Nos: 4 to 12) downstream of the sequence below, each of which
corresponds to the structure of any native human mRNA. Thus, a UNA
oligomer incorprating the oligomer fragment below can have enhanced
translational efficiency.
[0504] The translation enhancer is placed upstream of the AUG
translation start site, and the enhancer region is not translated
into the therapeutic protein.
TABLE-US-00021 (SEQ ID NO: 160) U{circumflex over (C)}AAC{tilde
over (A)}CAA{circumflex over (C)}AUA{tilde over (U)}ACAA{tilde over
(A)}AACAAAC{circumflex over (G)}AAU{circumflex over (C)}UCA{tilde
over (A)}GCA{tilde over (A)}UCA{tilde over (A)}GCA{tilde over
(U)}UC U{tilde over (A)}CUU{circumflex over (C)}UAU{tilde over
(U)}GCA{circumflex over (G)}CAA{tilde over (U)}UUA{tilde over
(A)}AU{circumflex over (C)}AUUU{circumflex over
(C)}UUUUAAA{circumflex over (G)}CAA{tilde over (A)}AGC{tilde over
(A)}AUU {tilde over (U)}UCU{circumflex over (G)}AAA{tilde over
(A)}UUU{tilde over (U)}CAC{circumflex over (C)}AUU{tilde over
(U)}ACG{tilde over (A)}ACG{tilde over (A)}UAGCC (SEQ ID NO: 161)
U{circumflex over (C)}AACACAACAUAUACAAAACAAACGAAUCU{circumflex over
(C)}AAGCAAUCAAGCAUUCU ACUUCUAUUGCA{circumflex over
(G)}CAAUUUAAAUCAUUUCUUUUAAAGCAAAA{circumflex over (G)}CAAUUU
UCUGAAAAUUUUCACCAUUUACGAACGAUAGC{circumflex over (C)}C (SEQ ID NO:
162) U{circumflex over (C)}{tilde over (A)}{tilde over
(A)}CACAACAUAUACAAAACAAACGAAUCUCAAGCAAUCAAGCAUUCU
ACUUCUAUUGCAGCAAUUUAAAUCAUUUCUUUUAAAGCAAAAGCAAUUU
UCUGAAAAUUUUCACCAUUUACGAACGAU{tilde over (A)}{circumflex over
(G)}{circumflex over (C)}CC (SEQ ID NO: 163) {tilde over
(U)}CAACACAACA{tilde over (U)}A{tilde over (U)}ACAAAACAAACGAA{tilde
over (U)}C{tilde over (U)}CAAGCAA{tilde over (U)}CAAGCA{tilde over
(U)}{tilde over (U)}C{tilde over (U)} AC{tilde over (U)}{tilde over
(U)}C{tilde over (U)}A{tilde over (U)}{tilde over (U)}GCAGCAA{tilde
over (U)}{tilde over (U)}{tilde over (U)}AAA{tilde over
(U)}CA{tilde over (U)}{tilde over (U)}{tilde over (U)}C{tilde over
(U)}{tilde over (U)}{tilde over (U)}{tilde over
(U)}AAAGCAAAAGCAA{tilde over (U)}{tilde over (U)}{tilde over (U)}
{tilde over (U)}C{tilde over (U)}GAAAA{tilde over (U)}{tilde over
(U)}{tilde over (U)}{tilde over (U)}CACCA{tilde over (U)}{tilde
over (U)}{tilde over (U)}ACGAACGA{tilde over (U)}AGCCC (SEQ ID NO:
164) {tilde over (U)}{circumflex over (C)}{tilde over (A)}{tilde
over (U)}{circumflex over (C)}{tilde over (A)}{circumflex over
(C)}{tilde over (A)}{tilde over (A)}{circumflex over (C)}{tilde
over (A)}{tilde over (U)}{tilde over (A)}{tilde over (U)}{tilde
over (A)}{circumflex over (C)}{tilde over (A)}{tilde over
(A)}{tilde over (A)}{tilde over (A)}{circumflex over (C)}{tilde
over (A)}{tilde over (A)}{tilde over (A)}{circumflex over
(C)}{circumflex over (G)}{tilde over (A)}{tilde over (A)}{tilde
over (U)}{circumflex over (C)}{tilde over (U)}{circumflex over
(C)}{tilde over (A)}{tilde over (A)}{circumflex over
(G)}{circumflex over (C)}{tilde over (A)}{tilde over (A)}{tilde
over (U)}{circumflex over (C)}{tilde over (A)}{tilde over
(A)}{circumflex over (G)}{circumflex over (C)}{tilde over
(A)}{tilde over (U)}{tilde over (U)}{circumflex over (C)}{tilde
over (U)} {tilde over (A)}{circumflex over (C)}{tilde over
(U)}{tilde over (U)}{circumflex over (C)}{tilde over (U)}{tilde
over (A)}{tilde over (U)}{tilde over (U)}{circumflex over
(G)}{circumflex over (C)}{tilde over (A)}{circumflex over
(G)}{circumflex over (C)}{tilde over (A)}{tilde over (A)}{tilde
over (U)}{tilde over (U)}{tilde over (U)}{tilde over (A)}{tilde
over (A)}{tilde over (A)}{tilde over (U)}{circumflex over
(C)}{tilde over (A)}{tilde over (U)}{tilde over (U)}{tilde over
(U)}{circumflex over (C)}{tilde over (U)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}{tilde over (A)}{tilde over (A)}{tilde
over (A)}{circumflex over (G)}{circumflex over (C)}{tilde over
(A)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{circumflex
over (G)}{circumflex over (C)}{tilde over (A)}{tilde over
(A)}{tilde over (U)}{tilde over (U)}{tilde over (U)} {tilde over
(U)}{circumflex over (C)}{tilde over (U)}{circumflex over
(G)}{tilde over (A)}{tilde over (A)}{tilde over (A)}{tilde over
(A)}{tilde over (U)}{tilde over (U)}{tilde over (U)}{tilde over
(U)}{circumflex over (C)}{tilde over (A)}{circumflex over
(C)}{circumflex over (C)}{tilde over (A)}{tilde over (U)}{tilde
over (U)}{tilde over (U)}{tilde over (A)}{circumflex over
(C)}{circumflex over (G)}{tilde over (A)}{tilde over
(A)}{circumflex over (C)}{circumflex over (G)}{tilde over
(A)}{tilde over (U)}{tilde over (A)}{circumflex over
(G)}{circumflex over (C)}{circumflex over (C)}{circumflex over
(C)}
Example 26
Messenger RNA Containing UNA Monomers
[0505] An nGFP transcript having a polyA tail of 30 monomers in
length is ligated to a donor poly tail of 30 UNA Monomers in length
to give an UNA-nGFP mRNA product having a .sub.polyA.sub.30 .sub.30
tail of 60 monomers in length. The UNA-nGFP has an increased
lifetime and markedly increased translational activity in
fibroblasts.
Example 27
Messenger RNA Containing UNA Monomers and Encoding HIV-1
Antigen
[0506] An mRNA encoding HIV-1 gag antigen having a polyA tail of 30
monomers in length is ligated to a donor poly tail of 20 UNA
Monomers in length to give an UNA-HIV-1 gag antigen mRNA product
having a polyA.sub.30 .sub.20 tail of 50 monomers in length. The
UNA-HIV-1 gag antigen mRNA has an increased lifetime and markedly
increased translational activity in fibroblasts.
Example 28
Messenger RNA Containing UNA Monomers and Encoding Lung Cancer
Antigens
[0507] An mRNA encoding antigens overexpressed in lung cancers
having a polyA tail of 30 monomers in length is ligated to a donor
poly tail of 10 UNA Monomers in length to give an UNA-mRNA product
having a polyA.sub.30 .sub.10 tail of 40 monomers in length. The
UNA-mRNA has an increased lifetime and markedly increased
translational activity in fibroblasts.
Example 29
Messenger RNA Containing UNA Monomers and Encoding Malarial P.
falciparum Reticulocyte-Binding Protein Homologue 5 (PfRH5)
[0508] An mRNA encoding malarial P. falciparum reticulocyte-binding
protein homologue 5 (PfRH5) having a polyA tail of 30 monomers in
length is ligated to a donor poly tail of 10 UNA Monomers in length
to give an UNA-mRNA product having a polyA.sub.30 .sub.10 tail of
40 monomers in length. The UNA-mRNA has an increased lifetime and
markedly increased translational activity in fibroblasts. The
UNA-mRNA is found to induce an antibody response in an animal
model.
Example 30
Messenger RNA Containing UNA Monomers and Encoding Malarial
Plasmodium falciparum PfSEA-1
[0509] An mRNA encoding malarial Plasmodium falciparum PfSEA-1, a
244 KD malaria antigen expressed in schizont-infected RBCs, having
a polyA tail of 30 monomers in length is ligated to a donor poly
tail of 10 UNA Monomers in length to give an UNA-mRNA product
having a polyA.sub.30 .sub.10 tail of 40 monomers in length. The
UNA-mRNA has an increased lifetime and markedly increased
translational activity in fibroblasts. The UNA-mRNA is found to
induce an antibody response in an animal model.
Example 31
Splint-Mediated Ligation
[0510] FIG. 7 shows the primary structure of a functional mRNA
transcript in the cytoplasm. The mRNA includes a 5' methylguanosine
cap, a protein coding sequence flanked by untranslated regions
(UTRs), and a polyadenosine (polyA) tail bound by polyA binding
proteins (PABPs).
[0511] FIG. 8 shows the 5' cap and PABPs cooperatively interacting
with proteins involved in translation to facilitate the recruitment
and recycling of ribosome complexes.
[0512] DNA splint oligomers were made for splint-mediated ligation
of of a donor oligomer to an acceptor RNA. As shown in the scheme
of FIG. 8, a short mRNA acceptor oligomer and a
5'-monophosphate-bearing polyA donor oligomer can be ligated in the
presence of a DNA splint oligomer.
[0513] FIG. 9 shows the splint-mediated ligation scheme, in which
an acceptor RNA with a 30-monomer stub polyA tail (A(30)) was
ligated to a 30-monomer donor oligomer (A(30)). The splint-mediated
ligation used a DNA oligomer splint which was complementary to the
3' UTR sequence upstream of the stub polyA tail, and included a
60-monomer oligo(dT) 5' heel (T(60)) to splint the ligation. The
anchoring region of the splint was complementary to the UTR
sequence to ensure that a 5' dT.sub.30 overhang was presented upon
hybridization to the acceptor. This brings the donor oligomer into
juxtaposition with the 3' terminus of the stub tail, dramatically
improving the kinetics of ligation.
[0514] FIG. 10 shows the results of ligation using 2 ug of a
120-monomer acceptor with an A.sub.30 stub tail that was ligated to
a 5'-phosphorylated A.sub.30 RNA donor oligomer using T4 RNA Ligase
2. The reaction was incubated overnight at 37.degree. C. The
ligation and a mock reaction done without enzyme were purified,
treated with DNAse I for 1 hour to degrade and detach the splint
oligomers, and re-purified in a volume of 30 uL. The ligation
efficiency was nearly 100%. The absence of a size shift in the
mock-reaction prep shows that the acceptor and donor were truly
ligated and not simply held together by undigested splint
oligomers.
[0515] Following the same protocol with a short incubation period,
high efficiency ligation of the short acceptor mRNA proceeded to
nearly 100% completion. FIG. 11 shows the results of
splint-mediated ligation using an acceptor RNA with a 30-monomer
stub polyA tail (A(30)). The ligation reactions were performed with
three different donor oligomer species: A(30), A(60), and A(120).
Based on the gel shifts, the ligations attained nearly 100%
efficiency.
Example 32
Splint-Mediated Ligation
[0516] A protocol used for a 100 ul splint-mediated ligation
reaction included the following materials, reagents, and steps.
[0517] 100 pmol UNA-PolyA UNA Oligomer donor.
[0518] 100 pmol TAIL-60 splint oligomer.
[0519] 50 pmol purified RNA acceptor.
[0520] 10 uL T4 RNA Ligase 2 10.times. Buffer.
[0521] 2 uL T4 RNA Ligase 2.
[0522] Nuclease-free Water to 100 uL.
[0523] Mix and incubate for 1-2 hours at 37 degrees, then purify
the RNA in a total of .about.90 uL RNAse-free water.
[0524] Add 10 uL 10.times. DNase buffer to eluent and 2 ul DNase I,
mix and incubate for 1 hour at 37 degrees to digest splint DNA.
[0525] Repurify the RNA using RNeasy spin columns, eluting in water
or TE pH 7.0.
[0526] Reagents.
[0527] NEB M0239 T4 RNA Ligase 2.
[0528] NEB M0303 DNase I (RNase-free).
[0529] Qiagen 74104RNeasy Mini Kit.
[0530] TAIL-60 splint oligomer sequence:
TABLE-US-00022 (SEQ ID NO: 165) CTTCCTACTCAGGCTTTATTCAAAGACCA.
[0531] Notes:
[0532] (a) The splint oligomer sequence includes an anchor that is
specific to the 3' UTR used for making mRNA.
[0533] (b) This protocol requires an mRNA transcript with a
pre-incorporated 30-nt polyA tail.
Example 33
Splint-Mediated Ligation
[0534] A full-length synthetic mRNA acceptor and a
5'-monophosphate-bearing polyA donor were ligated in the presence
of a DNA splint oligomer. On ligating a 30-monomer length tail to a
.about.1 Kb nGFP transcript, a size shift was apparent on a 2%
agarose gel, providing a direct indication that bulk ligation was
achieved. FIG. 12 shows the results of one-hour splint-mediated
ligations that were performed on nGFP-A.sub.30 transcripts. The
resulting ligation products were compared to untreated transcripts
and native nGFP-A.sub.60 IVT products. The native nGFP-A.sub.60 and
the ligated products were up-shifted on the gel relative to the
untreated nGFP-A.sub.30 transcripts and mock-ligated material.
Example 34
Splint-Mediated Ligation
[0535] A UNA-PolyA UNA Oligomer donor was made having the following
structure:
TABLE-US-00023 (SEQ ID NO: 166)
5'-(rAp)-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA -(3'-C3 Spacer),
wherein 5'-(rAp) is 5'-Phosphorylation and is UNA-A.
Example 35
Translatable RNA Molecules
[0536] An nGFP transcript with a polyA tail of 30-monomers in
length (untreated A.sub.30 mRNA) was ligated to a donor polyA tail
of 30-monomers in length to give an mRNA product having a polyA
tail of 60-monomers in length (A.sub.60-bearing ligation product)
by splint-mediated ligation.
[0537] FIG. 13 shows increased lifetime and translational activity
for the nGFP-A.sub.60 ligation product. As shown in FIG. 13,
nuclearized transcripts were transfected into fibroblasts for
comparison of nGFP-A.sub.30, mock-ligated nGFP-A.sub.30, and an
nGFP-A.sub.60 ligation product (FIG. 13, left to right). The
significantly higher fluorescence signal observed for the
nGFP-A.sub.60 ligation product shows that it has markedly increased
translational activity.
Example 36
Cohesive End Ligation
[0538] A wild-type T4 RNA ligase was used to ligate a donor 5'
phosphorylated oligomer to a short IVT transcript. Short synthetic
RNAs were generated by IVT, and the outcome of ligation reactions
was evaluated on high-resolution 4% agarose gels. The increase in
transcript size from ligation of a synthetic oligomer 30 monomers
in length to a full-sized mRNA of 1-2 Kb is too small to clearly
visualize on a gel. Thus, short synthetic RNAs of 100-180 monomers
were generated by IVT. The 3' terminal sequence of these short
synthetic RNAs was identical to that in the 3' UTRs of synthetic
mRNAs.
Example 37
Cohesive End Ligation with Pre-Adenylated Donor
[0539] A synthetic oligomer having an adenylated 5' end was
prepared. The adenylated 5' end, normally formed as a catalytic
intermediate by the ligase, pre-activated the synthetic oligomer
for ligation. Use of the pre-adenylated synthetic oligomer obviated
the need for ATP in the reactions, and allowed the use of a mutant
ligase that was active exclusively on adenylated substrates.
Pre-adenylation of the synthetic oligomer increased ligation
efficiency and minimized side-product formation.
[0540] A KQ mutant variant of T4 RNA Ligase 2 was used to ligate a
pre-adenylated donor oligomer to a short IVT transcript.
[0541] FIG. 14 shows the results of a ligation performed with a
100-monomer acceptor RNA that was treated for 3 hours at room
temperature with T4 RNA Ligase 2 (truncated KQ mutant) using a 10
uM concentration of a polyA tail 30-monomer donor oligomer. 15% PEG
8000 was included in the reaction as a volume excluder to promote
efficient ligation. The ligation reaction showed that a high
molecular weight product was formed, having a size in between the
100-monomer acceptor RNA and a 180-monomer RNA transcript included
as a size standard. These results show that the ligation reaction
produced a predominant product having high molecular weight with
nearly 100% ligation of the donor to the acceptor. Additional
experiments performed with concentrations of the polyA tail at 10
uM, 20 uM , and 40 uM showed that at least half of the acceptor RNA
was ligated in all cases.
[0542] All publications, patents and literature specifically
mentioned herein are incorporated by reference for all
purposes.
[0543] It is understood that this invention is not limited to the
particular methodology, protocols, materials, and reagents
described, as these may vary. It is also to be understood that the
terminology used herein is for the purpose of describing particular
embodiments only, and is not intended to limit the scope of the
present invention, which will be encompassed by the appended
claims.
[0544] It must be noted that as used herein and in the appended
claims, the singular forms "a", "an", and "the" include plural
reference unless the context clearly dictates otherwise. As well,
the terms "a" (or "an"), "one or more" and "at least one" can be
used interchangeably herein. It is also to be noted that the terms
"comprises," "comprising", "containing," "including", and "having"
can be used interchangeably.
[0545] Without further elaboration, it is believed that one skilled
in the art can, based on the above description, utilize the present
invention to its fullest extent. The following specific embodiments
are, therefore, to be construed as merely illustrative, and not
limitative of the remainder of the disclosure in any way
whatsoever.
[0546] All of the features disclosed in this specification may be
combined in any combination. Each feature disclosed in this
specification may be replaced by an alternative feature serving the
same, equivalent, or similar purpose.
Sequence CWU 1
1
16611750RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 1gggaaacaua agucaacaca acauauacaa
aacaaacgaa ucucaagcaa ucaagcauuc 60uacuucuauu gcagcaauuu aaaucauuuc
uuuuaaagca aaagcaauuu ucugaaaauu 120uucaccauuu acgaacgaua
gccauggccc agcgcgugaa caugaucaug gcagaaucac 180caggccucau
caccaucugc cuuuuaggau aucuacucag ugcugaaugu acaguuuuuc
240uugaucauga aaacgccaac aaaauucuga aucggccaaa gagguauaau
ucagguaaau 300uggaagaguu uguucaaggg aaccuugaga gagaauguau
ggaagaaaag uguaguuuug 360aagaagcacg agaaguuuuu gaaaacacug
aaagaacaac ugaauuuugg aagcaguaug 420uugauggaga ucagugugag
uccaauccau guuuaaaugg cggcaguugc aaggaugaca 480uuaauuccua
ugaauguugg ugucccuuug gauuugaagg aaagaacugu gaauuagaug
540uaacauguaa cauuaagaau ggcagaugcg agcaguuuug uaaaaauagu
gcugauaaca 600aggugguuug cuccuguacu gagggauauc gacuugcaga
aaaccagaag uccugugaac 660cagcagugcc auuuccaugu ggaagaguuu
cuguuucaca aacuucuaag cucacccgug 720cugagacugu uuuuccugau
guggacuaug uaaauucuac ugaagcugaa accauuuugg 780auaacaucac
ucaaagcacc caaucauuua augacuucac ucggguuguu gguggagaag
840augccaaacc aggucaauuc ccuuggcagg uuguuuugaa ugguaaaguu
gaugcauucu 900guggaggcuc uaucguuaau gaaaaaugga uuguaacugc
ugcccacugu guugaaacug 960guguuaaaau uacaguuguc gcaggugaac
auaauauuga ggagacagaa cauacagagc 1020aaaagcgaaa ugugauucga
auuauuccuc accacaacua caaugcagcu auuaauaagu 1080acaaccauga
cauugcccuu cuggaacugg acgaacccuu agugcuaaac agcuacguua
1140caccuauuug cauugcugac aaggaauaca cgaacaucuu ccucaaauuu
ggaucuggcu 1200auguaagugg cuggggaaga gucuuccaca aagggagauc
agcuuuaguu cuucaguacc 1260uuagaguucc acuuguugac cgagccacau
gucuucgauc uacaaaguuc accaucuaua 1320acaacauguu cugugcuggc
uuccaugaag gagguagaga uucaugucaa ggagauagug 1380ggggacccca
uguuacugaa guggaaggga ccaguuucuu aacuggaauu auuagcuggg
1440gugaagagug ugcaaugaaa ggcaaauaug gaauauauac caagguaucc
cgguauguca 1500acuggauuaa ggaaaaaaca aagcucacuu gacuagugac
ugacuaggau cugguuacca 1560cuaaaccagc cucaagaaca cccgaaugga
gucucuaagc uacauaauac caacuuacac 1620uuacaaaaug uuguccccca
aaauguagcc auucguaucu gcuccuaaua aaaagaaagu 1680uucuucacau
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
1740aaaaaaaaaa 17502943RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 2gggaaacaua agucaacaca
acauauacaa aacaaacgaa ucucaagcaa ucaagcauuc 60uacuucuauu gcagcaauuu
aaaucauuuc uuuuaaagca aaagcaauuu ucugaaaauu 120uucaccauuu
acgaacgaua gccauggggg ugcacgaaug uccugccugg cuguggcuuc
180uccugucccu gcugucgcuc ccucugggcc ucccaguccu gggcgcccca
ccacgccuca 240ucugugacag ccgaguccug gagagguacc ucuuggaggc
caaggaggcc gagaauauca 300cgacgggcug ugcugaacac ugcagcuuga
augagaauau cacuguccca gacaccaaag 360uuaauuucua ugccuggaag
aggauggagg ucgggcagca ggccguagaa gucuggcagg 420gccuggcccu
gcugucggaa gcuguccugc ggggccaggc ccuguugguc aacucuuccc
480agccguggga gccccugcag cugcaugugg auaaagccgu caguggccuu
cgcagccuca 540ccacucugcu ucgggcucug ggagcccaga aggaagccau
cuccccucca gaugcggccu 600cagcugcucc acuccgaaca aucacugcug
acacuuuccg caaacucuuc cgagucuacu 660ccaauuuccu ccggggaaag
cugaagcugu acacagggga ggccugcagg acaggggaca 720gaugacuagu
gacugacuag gaucugguua ccacuaaacc agccucaaga acacccgaau
780ggagucucua agcuacauaa uaccaacuua cacuuacaaa auguuguccc
ccaaaaugua 840gccauucgua ucugcuccua auaaaaagaa aguuucuuca
cauaaaaaaa aaaaaaaaaa 900aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaa 9433940DNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 3gggaaacaua
agucaacaca acauauacaa aacaaacgaa ucucaagcaa ucaagcauuc 60uacuucuauu
gcagcaauuu aaaucauuuc uuuuaaagca aaagcaauuu ucugaaaauu
120uucaccauuu acgaacgaua gccauggggg ugcccgaacg ucccacccug
cugcuuuuac 180ucuccuugcu acugauuccu cugggccucc caguccucug
ugcuccccca cgccucaucu 240gcgacagucg aguucuggag agguacaucu
uagaggccaa ggaggcagaa aaugucacga 300uggguugugc agaagguccc
agacugagug aaaauauuac agucccagau accaaaguca 360acuucuaugc
uuggaaaaga auggaggugg aagaacaggc cauagaaguu uggcaaggcc
420ugucccugcu cucagaagcc auccugcagg cccaggcccu gcuagccaau
uccucccagc 480caccagagac ccuucagcuu cauauagaca aagccaucag
uggucuacgu agccucacuu 540cacugcuucg gguacuggga gcucagaagg
aauugauguc gccuccagau accaccccac 600cugcuccacu ccgaacacuc
acaguggaua cuuucugcaa gcucuuccgg gucuacgcca 660acuuccuccg
ggggaaacug aagcuguaca cgggagaggu cugcaggaga ggggacaggt
720gacuagugac ugacuaggau cugguuacca cuaaaccagc cucaagaaca
cccgaaugga 780gucucuaagc uacauaauac caacuuacac uuacaaaaug
uuguccccca aaauguagcc 840auucguaucu gcuccuaaua aaaagaaagu
uucuucacau aaaaaaaaaa aaaaaaaaaa 900aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 940460RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 4aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
60560RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 5aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60660RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 6aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 60760RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 7aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
60860RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 8aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60960RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 9aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 601060RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 10aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
601160RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 11aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 601260RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 12aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 60131618RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 13gggaaacaua
agucaacaca acauauacaa aacaaacgaa ucucaagcaa ucaagcauuc 60uacuucuauu
gcagcaauuu aaaucauuuc uuuuaaagca aaagcaauuu ucugaaaauu
120uucaccauuu acgaacgaua gccaugccgu cuucugucuc guggggcauc
cuccugcugg 180caggccugug cugccugguc ccugucuccc uggcugagga
uccccaggga gaugcugccc 240agaagacaga uacaucccac caugaucagg
aucacccaac cuucaacaag aucaccccca 300accuggcuga guucgccuuc
agccuauacc gccagcuggc acaccagucc aacagcacca 360auaucuucuu
cuccccagug agcaucgcua cagccuuugc aaugcucucc cuggggacca
420aggcugacac ucacgaugaa auccuggagg gccugaauuu caaccucacg
gagauuccgg 480aggcucagau ccaugaaggc uuccaggaac uccuccguac
ccucaaccag ccagacagcc 540agcuccagcu gaccaccggc aauggccugu
uccucagcga gggccugaag cuaguggaua 600aguuuuugga ggauguuaaa
aaguuguacc acucagaagc cuucacuguc aacuucgggg 660acaccgaaga
ggccaagaaa cagaucaacg auuacgugga gaaggguacu caagggaaaa
720uuguggauuu ggucaaggag cuugacagag acacaguuuu ugcucuggug
aauuacaucu 780ucuuuaaagg caaaugggag agacccuuug aagucaagga
caccgaggaa gaggacuucc 840acguggacca ggugaccacc gugaaggugc
cuaugaugaa gcguuuaggc auguuuaaca 900uccagcacug uaagaagcug
uccagcuggg ugcugcugau gaaauaccug ggcaaugcca 960ccgccaucuu
cuuccugccu gaugagggga aacuacagca ccuggaaaau gaacucaccc
1020acgauaucau caccaaguuc cuggaaaaug aagacagaag gucugccagc
uuacauuuac 1080ccaaacuguc cauuacugga accuaugauc ugaagagcgu
ccugggucaa cugggcauca 1140cuaaggucuu cagcaauggg gcugaccucu
ccggggucac agaggaggca ccccugaagc 1200ucuccaaggc cgugcauaag
gcugugcuga ccaucgacga gaaagggacu gaagcugcug 1260gggccauguu
uuuagaggcc auacccaugu cuaucccccc cgaggucaag uucaacaaac
1320ccuuugucuu cuuaaugauu gaacaaaaua ccaagucucc ccucuucaug
ggaaaagugg 1380ugaaucccac ccaaaaauaa cuagugacug acuaggaucu
gguuaccacu aaaccagccu 1440caagaacacc cgaauggagu cucuaagcua
cauaauacca acuuacacuu acaaaauguu 1500gucccccaaa auguagccau
ucguaucugc uccuaauaaa aagaaaguuu cuucacauaa 1560aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa
161814943RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 14gggaaacaua agucaacaca acauauacaa
aacaaacgaa ucucaagcaa ucaagcauuc 60uacuucuauu gcagcaauuu aaaucauuuc
uuuuaaagca aaagcaauuu ucugaaaauu 120uucaccauuu acgaacgaua
gccauggggg ugcacgaaug uccugccugg cuguggcuuc 180uccugucccu
gcugucgcuc ccucugggcc ucccaguccu gggcgcccca ccacgccuca
240ucugugacag ccgaguccug gagagguacc ucuuggaggc caaggaggcc
gagaauauca 300cgacgggcug ugcugaacac ugcagcuuga augagaauau
cacuguccca gacaccaaag 360uuaauuucua ugccuggaag aggauggagg
ucgggcagca ggccguagaa gucuggcagg 420gccuggcccu gcugucggaa
gcuguccugc ggggccaggc ccuguugguc aacucuuccc 480agccguggga
gccccugcag cugcaugugg auaaagccgu caguggccuu cgcagccuca
540ccacucugcu ucgggcucug ggagcccaga aggaagccau cuccccucca
gaugcggccu 600cagcugcucc acuccgaaca aucacugcug acacuuuccg
caaacucuuc cgagucuacu 660ccaauuuccu ccggggaaag cugaagcugu
acacagggga ggccugcagg acaggggaca 720gaugacuagu gacugacuag
gaucugguua ccacuaaacc agccucaaga acacccgaau 780ggagucucua
agcuacauaa uaccaacuua cacuuacaaa auguuguccc ccaaaaugua
840gccauucgua ucugcuccua auaaaaagaa aguuucuuca cauaaaaaaa
aaaaaaaaaa 900aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa
943151386RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 15augcagcgcg ugaacaugau cauggcagaa
ucaccaggcc ucaucaccau cugccuuuua 60ggauaucuac ucagugcuga auguacaguu
uuucuugauc augaaaacgc caacaaaauu 120cugaaucggc caaagaggua
uaauucaggu aaauuggaag aguuuguuca agggaaccuu 180gagagagaau
guauggaaga aaaguguagu uuugaagaag cacgagaagu uuuugaaaac
240acugaaagaa caacugaauu uuggaagcag uauguugaug gagaucagug
ugaguccaau 300ccauguuuaa auggcggcag uugcaaggau gacauuaauu
ccuaugaaug uugguguccc 360uuuggauuug aaggaaagaa cugugaauua
gauguaacau guaacauuaa gaauggcaga 420ugcgagcagu uuuguaaaaa
uagugcugau aacaaggugg uuugcuccug uacugaggga 480uaucgacuug
cagaaaacca gaaguccugu gaaccagcag ugccauuucc auguggaaga
540guuucuguuu cacaaacuuc uaagcucacc cgugcugaga cuguuuuucc
ugauguggac 600uauguaaauu cuacugaagc ugaaaccauu uuggauaaca
ucacucaaag cacccaauca 660uuuaaugacu ucacucgggu uguuggugga
gaagaugcca aaccagguca auucccuugg 720cagguuguuu ugaaugguaa
aguugaugca uucuguggag gcucuaucgu uaaugaaaaa 780uggauuguaa
cugcugccca cuguguugaa acugguguua aaauuacagu ugucgcaggu
840gaacauaaua uugaggagac agaacauaca gagcaaaagc gaaaugugau
ucgaauuauu 900ccucaccaca acuacaaugc agcuauuaau aaguacaacc
augacauugc ccuucuggaa 960cuggacgaac ccuuagugcu aaacagcuac
guuacaccua uuugcauugc ugacaaggaa 1020uacacgaaca ucuuccucaa
auuuggaucu ggcuauguaa guggcugggg aagagucuuc 1080cacaaaggga
gaucagcuuu aguucuucag uaccuuagag uuccacuugu ugaccgagcc
1140acaugucuuc gaucuacaaa guucaccauc uauaacaaca uguucugugc
uggcuuccau 1200gaaggaggua gagauucaug ucaaggagau agugggggac
cccauguuac ugaaguggaa 1260gggaccaguu ucuuaacugg aauuauuagc
uggggugaag agugugcaau gaaaggcaaa 1320uauggaauau auaccaaggu
aucccgguau gucaacugga uuaaggaaaa aacaaagcuc 1380acuuaa
1386161386RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 16augcagcgcg ugaacaugau cauggcagaa
ucaccaggcc ucaucaccau cugccuuuua 60ggauaucuac ucagugcuga auguacaguu
uuucuugauc augaaaacgc caacaaaauu 120cugaaucggc caaagaggua
uaauucaggu aaauuggaag aguuuguuca agggaaccuu 180gagagagaau
guauggaaga aaaguguagu uuugaagaag cacgagaagu uuuugaaaac
240acugaaagaa caacugaauu uuggaagcag uauguugaug gagaucagug
ugaguccaau 300ccauguuuaa auggcggcag uugcaaggau gacauuaauu
ccuaugaaug uugguguccc 360uuuggauuug aaggaaagaa cugugaauua
gauguaacau guaacauuaa gaauggcaga 420ugcgagcagu uuuguaaaaa
uagugcugau aacaaggugg uuugcuccug uacugaggga 480uaucgacuug
cagaaaacca gaaguccugu gaaccagcag ugccauuucc auguggaaga
540guuucuguuu cacaaacuuc uaagcucacc cgugcugaga cuguuuuucc
ugauguggac 600uauguaaauu cuacugaagc ugaaaccauu uuggauaaca
ucacucaaag cacccaauca 660uuuaaugacu ucacucgggu uguuggugga
gaagaugcca aaccagguca auucccuugg 720cagguuguuu ugaaugguaa
aguugaugca uucuguggag gcucuaucgu uaaugaaaaa 780uggauuguaa
cugcugccca cuguguugaa acugguguua aaauuacagu ugucgcaggu
840gaacauaaua uugaggagac agaacauaca gagcaaaagc gaaaugugau
ucgaauuauu 900ccucaccaca acuacaaugc agcuauuaau aaguacaacc
augacauugc ccuucuggaa 960cuggacgaac ccuuagugcu aaacagcuac
guuacaccua uuugcauugc ugacaaggaa 1020uacacgaaca ucuuccucaa
auuuggaucu ggcuauguaa guggcugggg aagagucuuc 1080cacaaaggga
gaucagcuuu aguucuucag uaccuuagag uuccacuugu ugaccgagcc
1140acaugucuuc gaucuacaaa guucaccauc uauaacaaca uguucugugc
uggcuuccau 1200gaaggaggua gagauucaug ucaaggagau agugggggac
cccauguuac ugaaguggaa 1260gggaccaguu ucuuaacugg aauuauuagc
uggggugaag agugugcaau gaaaggcaaa 1320uauggaauau auaccaaggu
aucccgguau gucaacugga uuaaggaaaa aacaaagcuc 1380acuuaa
1386171386RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 17augcagcgcg ugaacaugau cauggcagaa
ucaccaggcc ucaucaccau cugccuuuua 60ggauaucuac ucagugcuga auguacaguu
uuucuugauc augaaaacgc caacaaaauu 120cugaaucggc caaagaggua
uaauucaggu aaauuggaag aguuuguuca agggaaccuu 180gagagagaau
guauggaaga aaaguguagu uuugaagaag cacgagaagu uuuugaaaac
240acugaaagaa caacugaauu uuggaagcag uauguugaug gagaucagug
ugaguccaau 300ccauguuuaa auggcggcag uugcaaggau gacauuaauu
ccuaugaaug uugguguccc 360uuuggauuug aaggaaagaa cugugaauua
gauguaacau guaacauuaa gaauggcaga 420ugcgagcagu uuuguaaaaa
uagugcugau aacaaggugg uuugcuccug uacugaggga 480uaucgacuug
cagaaaacca gaaguccugu gaaccagcag ugccauuucc auguggaaga
540guuucuguuu cacaaacuuc uaagcucacc cgugcugaga cuguuuuucc
ugauguggac 600uauguaaauu cuacugaagc ugaaaccauu uuggauaaca
ucacucaaag cacccaauca 660uuuaaugacu ucacucgggu uguuggugga
gaagaugcca aaccagguca auucccuugg 720cagguuguuu ugaaugguaa
aguugaugca uucuguggag gcucuaucgu uaaugaaaaa 780uggauuguaa
cugcugccca cuguguugaa acugguguua aaauuacagu ugucgcaggu
840gaacauaaua uugaggagac agaacauaca gagcaaaagc gaaaugugau
ucgaauuauu 900ccucaccaca acuacaaugc agcuauuaau aaguacaacc
augacauugc ccuucuggaa 960cuggacgaac ccuuagugcu aaacagcuac
guuacaccua uuugcauugc ugacaaggaa 1020uacacgaaca ucuuccucaa
auuuggaucu ggcuauguaa guggcugggg aagagucuuc 1080cacaaaggga
gaucagcuuu aguucuucag uaccuuagag uuccacuugu ugaccgagcc
1140acaugucuuc gaucuacaaa guucaccauc uauaacaaca uguucugugc
uggcuuccau 1200gaaggaggua gagauucaug ucaaggagau agugggggac
cccauguuac ugaaguggaa 1260gggaccaguu ucuuaacugg aauuauuagc
uggggugaag agugugcaau gaaaggcaaa 1320uauggaauau auaccaaggu
aucccgguau gucaacugga uuaaggaaaa aacaaagcuc 1380acuuaa
1386181257RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 18augccgucuu cugucucgug gggcauccuc
cugcuggcag gccugugcug ccuggucccu 60gucucccugg cugaggaucc ccagggagau
gcugcccaga agacagauac aucccaccau 120gaucaggauc acccaaccuu
caacaagauc acccccaacc uggcugaguu cgccuucagc 180cuauaccgcc
agcuggcaca ccaguccaac agcaccaaua ucuucuucuc cccagugagc
240aucgcuacag ccuuugcaau gcucucccug gggaccaagg cugacacuca
cgaugaaauc 300cuggagggcc ugaauuucaa ccucacggag auuccggagg
cucagaucca ugaaggcuuc 360caggaacucc uccguacccu caaccagcca
gacagccagc uccagcugac caccggcaau 420ggccuguucc ucagcgaggg
ccugaagcua guggauaagu uuuuggagga uguuaaaaag 480uuguaccacu
cagaagccuu cacugucaac uucggggaca ccgaagaggc caagaaacag
540aucaacgauu acguggagaa ggguacucaa gggaaaauug uggauuuggu
caaggagcuu 600gacagagaca caguuuuugc ucuggugaau uacaucuucu
uuaaaggcaa augggagaga 660cccuuugaag ucaaggacac cgaggaagag
gacuuccacg uggaccaggu gaccaccgug 720aaggugccua ugaugaagcg
uuuaggcaug uuuaacaucc agcacuguaa gaagcugucc 780agcugggugc
ugcugaugaa auaccugggc aaugccaccg ccaucuucuu ccugccugau
840gaggggaaac uacagcaccu ggaaaaugaa cucacccacg auaucaucac
caaguuccug 900gaaaaugaag acagaagguc ugccagcuua cauuuaccca
aacuguccau uacuggaacc 960uaugaucuga agagcguccu gggucaacug
ggcaucacua aggucuucag caauggggcu 1020gaccucuccg gggucacaga
ggaggcaccc cugaagcucu ccaaggccgu gcauaaggcu 1080gugcugacca
ucgacgagaa agggacugaa gcugcugggg ccauguuuuu agaggccaua
1140cccaugucua ucccccccga ggucaaguuc aacaaacccu uugucuucuu
aaugauugaa 1200caaaauacca agucuccccu cuucauggga aaagugguga
aucccaccca aaaauaa 1257191257RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 19augccgucuu
cugucucgug gggcauccuc cugcuggcag gccugugcug ccuggucccu 60gucucccugg
cugaggaucc ccagggagau gcugcccaga agacagauac aucccaccau
120gaucaggauc acccaaccuu caacaagauc acccccaacc uggcugaguu
cgccuucagc 180cuauaccgcc agcuggcaca ccaguccaac agcaccaaua
ucuucuucuc cccagugagc 240aucgcuacag ccuuugcaau gcucucccug
gggaccaagg cugacacuca cgaugaaauc 300cuggagggcc ugaauuucaa
ccucacggag auuccggagg cucagaucca ugaaggcuuc 360caggaacucc
uccguacccu caaccagcca gacagccagc uccagcugac caccggcaau
420ggccuguucc ucagcgaggg ccugaagcua guggauaagu uuuuggagga
uguuaaaaag 480uuguaccacu cagaagccuu cacugucaac uucggggaca
ccgaagaggc caagaaacag 540aucaacgauu acguggagaa ggguacucaa
gggaaaauug uggauuuggu caaggagcuu 600gacagagaca caguuuuugc
ucuggugaau uacaucuucu uuaaaggcaa augggagaga 660cccuuugaag
ucaaggacac cgaggaagag gacuuccacg uggaccaggu gaccaccgug
720aaggugccua ugaugaagcg uuuaggcaug uuuaacaucc agcacuguaa
gaagcugucc 780agcugggugc ugcugaugaa auaccugggc aaugccaccg
ccaucuucuu ccugccugau 840gaggggaaac uacagcaccu ggaaaaugaa
cucacccacg auaucaucac caaguuccug 900gaaaaugaag acagaagguc
ugccagcuua cauuuaccca aacuguccau uacuggaacc 960uaugaucuga
agagcguccu gggucaacug ggcaucacua aggucuucag caauggggcu
1020gaccucuccg gggucacaga ggaggcaccc
cugaagcucu ccaaggccgu gcauaaggcu 1080gugcugacca ucgacgagaa
agggacugaa gcugcugggg ccauguuuuu agaggccaua 1140cccaugucua
ucccccccga ggucaaguuc aacaaacccu uugucuucuu aaugauugaa
1200caaaauacca agucuccccu cuucauggga aaagugguga aucccaccca aaaauaa
1257201257RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 20augccgucuu cugucucgug gggcauccuc
cugcuggcag gccugugcug ccuggucccu 60gucucccugg cugaggaucc ccagggagau
gcugcccaga agacagauac aucccaccau 120gaucaggauc acccaaccuu
caacaagauc acccccaacc uggcugaguu cgccuucagc 180cuauaccgcc
agcuggcaca ccaguccaac agcaccaaua ucuucuucuc cccagugagc
240aucgcuacag ccuuugcaau gcucucccug gggaccaagg cugacacuca
cgaugaaauc 300cuggagggcc ugaauuucaa ccucacggag auuccggagg
cucagaucca ugaaggcuuc 360caggaacucc uccguacccu caaccagcca
gacagccagc uccagcugac caccggcaau 420ggccuguucc ucagcgaggg
ccugaagcua guggauaagu uuuuggagga uguuaaaaag 480uuguaccacu
cagaagccuu cacugucaac uucggggaca ccgaagaggc caagaaacag
540aucaacgauu acguggagaa ggguacucaa gggaaaauug uggauuuggu
caaggagcuu 600gacagagaca caguuuuugc ucuggugaau uacaucuucu
uuaaaggcaa augggagaga 660cccuuugaag ucaaggacac cgaggaagag
gacuuccacg uggaccaggu gaccaccgug 720aaggugccua ugaugaagcg
uuuaggcaug uuuaacaucc agcacuguaa gaagcugucc 780agcugggugc
ugcugaugaa auaccugggc aaugccaccg ccaucuucuu ccugccugau
840gaggggaaac uacagcaccu ggaaaaugaa cucacccacg auaucaucac
caaguuccug 900gaaaaugaag acagaagguc ugccagcuua cauuuaccca
aacuguccau uacuggaacc 960uaugaucuga agagcguccu gggucaacug
ggcaucacua aggucuucag caauggggcu 1020gaccucuccg gggucacaga
ggaggcaccc cugaagcucu ccaaggccgu gcauaaggcu 1080gugcugacca
ucgacgagaa agggacugaa gcugcugggg ccauguuuuu agaggccaua
1140cccaugucua ucccccccga ggucaaguuc aacaaacccu uugucuucuu
aaugauugaa 1200caaaauacca agucuccccu cuucauggga aaagugguga
aucccaccca aaaauaa 12572141RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 21ggcaccacca
cugaccuggg acagugaauc gacagccgac c 412241RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 22ggcaccacca cugaccuggg acagugaauc gacagccgac c
412341RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 23ggcaccacca cugaccuggg acagugaauc
gacagccgac c 412441RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 24ggcaccacca cugaccuggg
acagugaauc gacagccgac c 412541RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 25ggcaccacca
cugaccuggg acagugaauc gacagccgac c 412641RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 26ggcaccacca cugaccuggg acagugaauc gacagccgac c
412741RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 27ggcaccacca cugaccuggg acagugaauc
gacagccgac c 412841RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 28ggcaccacca cugaccuggg
acagugaauc gacagccgac c 412941RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 29ggcaccacca
cugaccuggg acagugaauc gacagccgac c 413041RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 30ggcaccacca cugaccuggg acagugaauc gacagccgac c
413141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 31ggcaccacca cugaccuggg acagugaauc
gacagccgac c 413241RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 32ggcaccacca cugaccuggg
acagugaauc gacagccgac c 413341RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 33ggcaccacca
cugaccuggg acagugaauc gacagccgac c 413441RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 34ggcaccacca cugaccuggg acagugaauc gacagccgac c
413541RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 35ggcaccacca cugaccuggg acagugaauc
gacagccgac c 413641RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 36ggcaccacca cugaccuggg
acagugaauc gacagccgac c 413741RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 37ggcaccacca
cugaccuggg acagugaauc gacagccgac c 413841RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 38ggcaccacca cugaccuggg acagugaauc gacagccgac c
413941RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 39ggcaccacca cugaccuggg acagugaauc
gacagccgac c 414041RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 40ggcaccacca cugaccuggg
acagugaauc gacagccgac c 414141RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 41ggcaccacca
cugaccuggg acagugaauc gacagccgac c 414241RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 42ggcaccacca cugaccuggg acagugaauc gacagccgac c
414341RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 43ggcaccacca cugaccuggg acagugaauc
gacagccgac c 414441RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 44ggcaccacca cugaccuggg
acagugaauc gacagccgac c 414541RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 45ggcaccacca
cugaccuggg acagugaauc gacagccgac c 414641RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 46ggcaccacca cugaccuggg acagugaauc gacagccgac c
414741RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 47ggcaccacca cugaccuggg acagugaauc
gacagccgac c 414841RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 48ggcaccacca cugaccuggg
acagugaauc gacagccgac c 414941RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 49ggcaccacca
cugaccuggg acagugaauc gacagccgac c 415041RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 50ggcaccacca cugaccuggg acagugaauc gacagccgac c
415141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 51ggcaccacca cugaccuggg acagugaauc
gacagccgac c 415241RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 52ggcaccacca cugaccuggg
acagugaauc gacagccgac c 415341RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 53ggcaccacca
cugaccuggg acagugaauc gacagccgac c 415441RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 54ggcaccacca cugaccuggg acagugaauc gacagccgac c
415541RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 55ggcaccacca cugaccuggg acagugaauc
gacagccgac c 415641RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 56ggcaccacca cugaccuggg
acagugaauc gacagccgac c 415741RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 57ggcaccacca
cugaccuggg acagugaauc gacagccgac c 415841RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 58ggcaccacca cugaccuggg acagugaauc gacagccgac c
415941RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 59ggcaccacca cugaccuggg acagugaauc
gacagccgac c 416041RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 60ggcaccacca cugaccuggg
acagugaauc gacagccgac c 416141RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 61ggcaccacca
cugaccuggg acagugaauc gacagccgac c 416241RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 62ggcaccacca cugaccuggg acagugaauc gacagccgac c
416341RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 63ggcaccacca cugaccuggg acagugaauc
gacagccgac c 416441RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 64ggcaccacca cugaccuggg
acagugaauc gacagccgac c 416541RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 65ggcaccacca
cugaccuggg acagugaauc gacagccgac c 416641RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 66ggcaccacca cugaccuggg acagugaauc gacagccgac c
416741RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 67ggcaccacca cugaccuggg acagugaauc
gacagccgac c 416841RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 68ggcaccacca cugaccuggg
acagugaauc gacagccgac c 416941RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 69ggcaccacca
cugaccuggg acagugaauc gacagccgac c 417041RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 70ggcaccacca cugaccuggg acagugaauc gacagccgac c
417141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 71ggcaccacca cugaccuggg acagugaauc
gacagccgac c 417241RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 72ggcaccacca cugaccuggg
acagugaauc gacagccgac c 417341RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 73ggcaccacca
cugaccuggg acagugaauc gacagccgac c 417441RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 74ggcaccacca cugaccuggg acagugaauc gacagccgac c
417541RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 75ggcaccacca cugaccuggg acagugaauc
gacagccgac c 417641RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 76ggcaccacca cugaccuggg
acagugaauc gacagccgac c 417741RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 77ggcaccacca
cugaccuggg acagugaauc gacagccgac c 417841RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 78ggcaccacca cugaccuggg acagugaauc gacagccgac c
417941RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 79ggcaccacca cugaccuggg acagugaauc
gacagccgac c 418041RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 80ggcaccacca cugaccuggg
acagugaauc gacagccgac c 418141RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 81ggcaccacca
cugaccuggg acagugaauc gacagccgac c 418241RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 82ggcaccacca cugaccuggg acagugaauc gacagccgac c
418341RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 83ggcaccacca cugaccuggg acagugaauc
gacagccgac c 418441RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 84ggcaccacca cugaccuggg
acagugaauc gacagccgac c 418541RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 85ggcaccacca
cugaccuggg acagugaauc gacagccgac c 418641RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 86ggcaccacca cugaccuggg acagugaauc gacagccgac c
418741RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 87ggcaccacca cugaccuggg acagugaauc
gacagccgac c 418841RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 88ggcaccacca cugaccuggg
acagugaauc gacagccgac c 418941RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 89ggcaccacca
cugaccuggg acagugaauc gacagccgac c 419041RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 90ggcaccacca cugaccuggg acagugaauc gacagccgac c
419141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 91ggcaccacca cugaccuggg acagugaauc
gacagccgac c 419241RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 92ggcaccacca cugaccuggg
acagugaauc gacagccgac c 419341RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 93ggcaccacca
cugaccuggg acagugaauc gacagccgac c 419441RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 94ggcaccacca cugaccuggg acagugaauc gacagccgac c
419541RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 95ggcaccacca cugaccuggg acagugaauc
gacagccgac c 419641RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 96ggcaccacca cugaccuggg
acagugaauc gacagccgac c 419741RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 97ggcaccacca
cugaccuggg acagugaauc gacagccgac c 419841RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 98ggcaccacca cugaccuggg acagugaauc gacagccgac c
419941RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 99ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4110041RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 100ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4110141RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 101ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4110241RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 102ggcaccacca cugaccuggg acagugaauc gacagccgac c
4110341RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 103ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4110441RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 104ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4110541RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 105ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4110641RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 106ggcaccacca cugaccuggg acagugaauc gacagccgac c
4110741RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 107ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4110841RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 108ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4110941RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 109ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4111041RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 110ggcaccacca cugaccuggg acagugaauc gacagccgac c
4111141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 111ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4111241RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 112ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4111341RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 113ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4111441RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 114ggcaccacca cugaccuggg acagugaauc gacagccgac c
4111541RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 115ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4111641RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 116ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4111741RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 117ggcaccacca cugaccuggg acagugaauc gacagccgac c
4111841RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 118ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4111941RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 119ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4112041RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 120ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4112141RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 121ggcaccacca cugaccuggg acagugaauc gacagccgac c
4112241RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 122ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4112341RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 123ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4112441RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 124ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4112541RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 125ggcaccacca cugaccuggg acagugaauc gacagccgac c
4112641RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 126ggcaccacca cugaccuggg acagugaauc
gacagccgac c 4112741RNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 127ggcaccacca cugaccuggg
acagugaauc gacagccgac c 4112841RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 128ggcaccacca
cugaccuggg acagugaauc gacagccgac c 4112941RNAArtificial
SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 129ggcaccacca cugaccuggg acagugaauc gacagccgac c
4113041RNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 130ggcaccacca cugaccuggg acagugaauc
gacagccgac c 41131582RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 131augggggugc
acgaaugucc ugccuggcug uggcuucucc ugucccugcu gucgcucccu 60cugggccucc
caguccuggg cgccccacca cgccucaucu gugacagccg aguccuggag
120agguaccucu uggaggccaa ggaggccgag aauaucacga cgggcugugc
ugaacacugc 180agcuugaaug agaauaucac ugucccagac accaaaguua
auuucuaugc cuggaagagg 240auggaggucg ggcagcaggc cguagaaguc
uggcagggcc uggcccugcu gucggaagcu 300guccugcggg gccaggcccu
guuggucaac ucuucccagc cgugggagcc ccugcagcug 360cauguggaua
aagccgucag uggccuucgc agccucacca cucugcuucg ggcucuggga
420gcccagaagg aagccaucuc cccuccagau gcggccucag cugcuccacu
ccgaacaauc 480acugcugaca cuuuccgcaa acucuuccga gucuacucca
auuuccuccg gggaaagcug 540aagcuguaca caggggaggc cugcaggaca
ggggacagau ga 582132582RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 132augggggugc
acgaaugucc ugccuggcug uggcuucucc ugucccugcu gucgcucccu 60cugggccucc
caguccuggg cgccccacca cgccucaucu gugacagccg aguccuggag
120agguaccucu uggaggccaa ggaggccgag aauaucacga cgggcugugc
ugaacacugc 180agcuugaaug agaauaucac ugucccagac accaaaguua
auuucuaugc cuggaagagg 240auggaggucg ggcagcaggc cguagaaguc
uggcagggcc uggcccugcu gucggaagcu 300guccugcggg gccaggcccu
guuggucaac ucuucccagc cgugggagcc ccugcagcug 360cauguggaua
aagccgucag uggccuucgc agccucacca cucugcuucg ggcucuggga
420gcccagaagg aagccaucuc cccuccagau gcggccucag cugcuccacu
ccgaacaauc 480acugcugaca cuuuccgcaa acucuuccga gucuacucca
auuuccuccg gggaaagcug 540aagcuguaca caggggaggc cugcaggaca
ggggacagau ga 582133582RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 133augggggugc
acgaaugucc ugccuggcug uggcuucucc ugucccugcu gucgcucccu 60cugggccucc
caguccuggg cgccccacca cgccucaucu gugacagccg aguccuggag
120agguaccucu uggaggccaa ggaggccgag aauaucacga cgggcugugc
ugaacacugc 180agcuugaaug agaauaucac ugucccagac accaaaguua
auuucuaugc cuggaagagg 240auggaggucg ggcagcaggc cguagaaguc
uggcagggcc uggcccugcu gucggaagcu 300guccugcggg gccaggcccu
guuggucaac ucuucccagc cgugggagcc ccugcagcug 360cauguggaua
aagccgucag uggccuucgc agccucacca cucugcuucg ggcucuggga
420gcccagaagg aagccaucuc cccuccagau gcggccucag cugcuccacu
ccgaacaauc 480acugcugaca cuuuccgcaa acucuuccga gucuacucca
auuuccuccg gggaaagcug 540aagcuguaca caggggaggc cugcaggaca
ggggacagau ga 5821341066RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 134augcuguuua
aucugaggau ccuguuuaaa caaugcagcu uuuagaaaug gucacaacuu 60caugguucga
aauuuucggu guggacaacc acuacaaaau aaagugcagc ugaagggccg
120ugaccuucuc acucuaaaaa acuuuaccgg agaagaaauu aaauauaugc
uauggcuauc 180agcagaucug aaauuuagga uaaaacagaa aggagaguau
uugccuuuau ugcaagggaa 240guccuuaggc augauuuuug agaaaagaag
uacucgaaca agauugucua cagaaacagg 300cuuugcacuu cugggaggac
auccuuguuu ucuuaccaca caagauauuc auuugggugu 360gaaugaaagu
cucacggaca cggcccgugu auugucuagc auggcagaug caguauuggc
420ucgaguguau aaacaaucag auuuggacac ccuggcuaaa gaagcaucca
ucccaauuau 480caaugggcug ucagauuugu accauccuau ccagauccug
gcugauuacc ucacgcucca 540ggaacacuau agcucucuga aaggucuuac
ccucagcugg aucggggaug ggaacaauau 600ccugcacucc aucaugauga
gcgcagcgaa auucggaaug caccuucagg cagcuacucc 660aaaggguuau
gagccggaug cuaguguaac caaguuggca gagcaguaug ccaaagagaa
720ugguaccaag cuguugcuga caaaugaucc auuggaagca gcgcauggag
gcaauguauu 780aauuacagac acuuggauaa gcaugggaca agaagaggag
aagaaaaagc ggcuccaggc 840uuuccaaggu uaccagguua caaugaagac
ugcuaaaguu gcugccucug acuggacauu 900uuuacacugc uugcccagaa
agccagaaga aguggaugau gaagucuuuu auucuccucg 960aucacuagug
uucccagagg cagaaaacag aaaguggaca aucauggcug ucaugguguc
1020ccugcugaca gauuacucac cucagcucca gaagccuaaa uuuuga
10661351065RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 135augcuguuua aucugaggau ccuguuaaac
aaugcagcuu uuagaaaugg ucacaacuuc 60augguucgaa auuuucggug uggacaacca
cuacaaaaua aagugcagcu gaagggccgu 120gaccuucuca cucuaaaaaa
cuuuaccgga gaagaaauua aauauaugcu auggcuauca 180gcagaucuga
aauuuaggau aaaacagaaa ggagaguauu ugccuuuauu gcaagggaag
240uccuuaggca ugauuuuuga gaaaagaagu acucgaacaa gauugucuac
agaaacaggc 300uuugcacuuc ugggaggaca uccuuguuuu cuuaccacac
aagauauuca uuugggugug 360aaugaaaguc ucacggacac ggcccgugua
uugucuagca uggcagaugc aguauuggcu 420cgaguguaua aacaaucaga
uuuggacacc cuggcuaaag aagcauccau cccaauuauc 480aaugggcugu
cagauuugua ccauccuauc cagauccugg cugauuaccu cacgcuccag
540gaacacuaua gcucucugaa aggucuuacc cucagcugga ucggggaugg
gaacaauauc 600cugcacucca ucaugaugag cgcagcgaaa uucggaaugc
accuucaggc agcuacucca 660aaggguuaug agccggaugc uaguguaacc
aaguuggcag agcaguaugc caaagagaau 720gguaccaagc uguugcugac
aaaugaucca uuggaagcag cgcauggagg caauguauua 780auuacagaca
cuuggauaag caugggacaa gaagaggaga agaaaaagcg gcuccaggcu
840uuccaagguu accagguuac aaugaagacu gcuaaaguug cugccucuga
cuggacauuu 900uuacacugcu ugcccagaaa gccagaagaa guggaugaug
aagucuuuua uucuccucga 960ucacuagugu ucccagaggc agaaaacaga
aaguggacaa ucauggcugu cauggugucc 1020cugcugacag auuacucacc
ucagcuccag aagccuaaau uuuga 10651361065RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
136augcuguuua aucugaggau ccuguuaaac aaugcagcuu uuagaaaugg
ucacaacuuc 60augguucgaa auuuucggug uggacaacca cuacaaaaua aagugcagcu
gaagggccgu 120gaccuucuca cucuaaaaaa cuuuaccgga gaagaaauua
aauauaugcu auggcuauca 180gcagaucuga aauuuaggau aaaacagaaa
ggagaguauu ugccuuuauu gcaagggaag 240uccuuaggca ugauuuuuga
gaaaagaagu acucgaacaa gauugucuac agaaacaggc 300uuugcacuuc
ugggaggaca uccuuguuuu cuuaccacac aagauauuca uuugggugug
360aaugaaaguc ucacggacac ggcccgugua uugucuagca uggcagaugc
aguauuggcu 420cgaguguaua aacaaucaga uuuggacacc cuggcuaaag
aagcauccau cccaauuauc 480aaugggcugu cagauuugua ccauccuauc
cagauccugg cugauuaccu cacgcuccag 540gaacacuaua gcucucugaa
aggucuuacc cucagcugga ucggggaugg gaacaauauc 600cugcacucca
ucaugaugag cgcagcgaaa uucggaaugc accuucaggc agcuacucca
660aaggguuaug agccggaugc uaguguaacc aaguuggcag agcaguaugc
caaagagaau 720gguaccaagc uguugcugac aaaugaucca uuggaagcag
cgcauggagg caauguauua 780auuacagaca cuuggauaag caugggacaa
gaagaggaga agaaaaagcg gcuccaggcu 840uuccaagguu accagguuac
aaugaagacu gcuaaaguug cugccucuga cuggacauuu 900uuacacugcu
ugcccagaaa gccagaagaa guggaugaug aagucuuuua uucuccucga
960ucacuagugu ucccagaggc agaaaacaga aaguggacaa ucauggcugu
cauggugucc 1020cugcugacag auuacucacc ucagcuccag aagccuaaau uuuga
1065137134RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 137gcucgcuuuc uugcugucca auuucuauua
aagguuccuu uguucccuaa guccaacuac 60uaaacugggg gauauuauga agggccuuga
gcaucuggau ucugccuaau aaaaaacauu 120uauuuucauu gcaa
134138134RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 138gcucgcuuuc uugcugucca auuucuauua
aagguuccuu uguucccuaa guccaacuac 60uaaacugggg gauauuauga agggccuuga
gcaucuggau ucugccuaau aaaaaacauu 120uauuuucauu gcaa
134139134RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 139gcucgcuuuc uugcugucca auuucuauua
aagguuccuu uguucccuaa guccaacuac 60uaaacugggg gauauuauga agggccuuga
gcaucuggau ucugccuaau aaaaaacauu 120uauuuucauu gcaa
134140134RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 140gcucgcuuuc uugcugucca auuucuauua
aagguuccuu uguucccuaa guccaacuac 60uaaacugggg gauauuauga agggccuuga
gcaucuggau ucugccuaau aaaaaacauu 120uauuuucauu gcaa
134141134RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 141gcucgcuuuc uugcugucca auuucuauua
aagguuccuu uguucccuaa guccaacuac 60uaaacugggg gauauuauga agggccuuga
gcaucuggau ucugccuaau aaaaaacauu 120uauuuucauu gcaa
134142158RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 142cuagugacug acuaggaucu gguuaccacu
aaaccagccu caagaacacc cgaauggagu 60cucuaagcua cauaauacca acuuacacuu
acaaaauguu gucccccaaa auguagccau 120ucguaucugc uccuaauaaa
aagaaaguuu cuucacau 158143158RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 143cuagugacug
acuaggaucu gguuaccacu aaaccagccu caagaacacc cgaauggagu 60cucuaagcua
cauaauacca acuuacacuu acaaaauguu gucccccaaa auguagccau
120ucguaucugc uccuaauaaa aagaaaguuu cuucacau 158144158RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
144cuagugacug acuaggaucu gguuaccacu aaaccagccu caagaacacc
cgaauggagu 60cucuaagcua cauaauacca acuuacacuu acaaaauguu gucccccaaa
auguagccau 120ucguaucugc uccuaauaaa aagaaaguuu cuucacau
158145158RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 145cuagugacug acuaggaucu gguuaccacu
aaaccagccu caagaacacc cgaauggagu 60cucuaagcua cauaauacca acuuacacuu
acaaaauguu gucccccaaa auguagccau 120ucguaucugc uccuaauaaa
aagaaaguuu cuucacau 158146158RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 146cuagugacug
acuaggaucu gguuaccacu aaaccagccu caagaacacc cgaauggagu 60cucuaagcua
cauaauacca acuuacacuu acaaaauguu gucccccaaa auguagccau
120ucguaucugc uccuaauaaa aagaaaguuu cuucacau
1581471063RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 147auggagcuga cugaauugcu ccucgugguc
augcuucucc uaacugcaag gcuaacgcug 60uccagcccgg cuccuccugc uugugaccuc
cgaguccuca guaaacugcu ucgugacucc 120cauguccuuc acagcagacu
gagccagugc ccagagguuc acccuuugcc uacaccuguc 180cugcugccug
cuguggacuu uagcuuggga gaauggaaaa cccagaugga ggagaccaag
240gcacaggaca uucugggagc agugacccuu cugcuggagg gagugauggc
agcacgggga 300caacugggac ccacuugccu cucaucccuc cuggggcagc
uuucuggaca gguccgucuc 360cuccuugggg cccugcagag ccuccuugga
acccagcuuc cuccacaggg caggaccaca 420gcucacaagg aucccaaugc
caucuuccug agcuuccaac accugcuccg aggaaaggug 480cguuuccuga
ugcuuguagg aggguccacc cucugcguca ggcggggccc cacccaccac
540agcugucccc agcagaaccu cucuaguccu cacacugaac gagcucccaa
acaggacuuc 600uggauuguug gagacaaacu ucacugccuc agccagaacu
acuggcucug ggcuucugaa 660guggcagcag ggauucagag ccaagauucc
uggucugcug aaccaaaccu ccaggucccu 720ggaccaaauc cccggauacc
ugaacaggau acacgaacuc uugaauggaa cucguggacu 780cuuuccugga
cccucacgca ggacccuagg agccccggac auuuccucag gaacaucaga
840cacaggcucc cugccaccca accuccagcc uggauauucu ccuuccccaa
cccauccucc 900uacuggacag uauacgcucu ucccucuucc acccaccuug
cccaccccug ugguccagcu 960ccacccccug cuuccugacc cuucugcucc
aacgcccacc ccuaccagcc cucuucuaaa 1020cacauccuac acccacuccc
agaaucuguc ucaggaaggg uaa 10631481062RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
148auggagcuga cugaauugcu ccucgugguc augcuucucc uaacugcaag
gcuaacgcug 60uccagcccgg cuccuccugc uugugaccuc cgaguccuca guaaacugcu
ucgugacucc 120cauguccuuc acagcagacu gagccagugc ccagagguuc
acccuuugcc uacaccuguc 180cugcugccug cuguggacuu uagcuuggga
gaauggaaaa cccagaugga ggagaccaag 240gcacaggaca uucugggagc
agugacccuu cugcuggagg gagugauggc agcacgggga 300caacugggac
ccacuugccu cucaucccuc cuggggcagc uuucuggaca gguccgucuc
360cuccuugggg cccugcagag ccuccuugga acccagcuuc cuccacaggg
caggaccaca 420gcucacaagg aucccaaugc caucuuccug agcuuccaac
accugcuccg aggaaaggug 480cguuuccuga ugcuuguagg aggguccacc
cucugcguca ggcgggcccc acccaccaca 540gcugucccca gcagaaccuc
ucuaguccuc acacugaacg agcucccaaa caggacuucu 600ggauuguugg
agacaaacuu cacugccuca gccagaacua cuggcucugg gcuucugaag
660uggcagcagg gauucagagc caagauuccu ggucugcuga accaaaccuc
caggucccug 720gaccaaaucc ccggauaccu gaacaggaua cacgaacucu
ugaauggaac ucguggacuc 780uuuccuggac ccucacgcag gacccuagga
gccccggaca uuuccucagg aacaucagac 840acaggcuccc ugccacccaa
ccuccagccu ggauauucuc cuuccccaac ccauccuccu 900acuggacagu
auacgcucuu cccucuucca cccaccuugc ccaccccugu gguccagcuc
960cacccccugc uuccugaccc uucugcucca acgcccaccc cuaccagccc
ucuucuaaac 1020acauccuaca cccacuccca gaaucugucu caggaagggu aa
10621491062RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 149auggagcuga cugaauugcu ccucgugguc
augcuucucc uaacugcaag gcuaacgcug 60uccagcccgg cuccuccugc uugugaccuc
cgaguccuca guaaacugcu ucgugacucc 120cauguccuuc acagcagacu
gagccagugc ccagagguuc acccuuugcc uacaccuguc 180cugcugccug
cuguggacuu uagcuuggga gaauggaaaa cccagaugga ggagaccaag
240gcacaggaca uucugggagc agugacccuu cugcuggagg gagugauggc
agcacgggga 300caacugggac ccacuugccu cucaucccuc cuggggcagc
uuucuggaca gguccgucuc 360cuccuugggg cccugcagag ccuccuugga
acccagcuuc cuccacaggg caggaccaca 420gcucacaagg aucccaaugc
caucuuccug agcuuccaac accugcuccg aggaaaggug 480cguuuccuga
ugcuuguagg aggguccacc cucugcguca ggcgggcccc acccaccaca
540gcugucccca gcagaaccuc ucuaguccuc acacugaacg agcucccaaa
caggacuucu 600ggauuguugg agacaaacuu cacugccuca gccagaacua
cuggcucugg gcuucugaag 660uggcagcagg gauucagagc caagauuccu
ggucugcuga accaaaccuc caggucccug 720gaccaaaucc ccggauaccu
gaacaggaua cacgaacucu ugaauggaac ucguggacuc 780uuuccuggac
ccucacgcag gacccuagga gccccggaca uuuccucagg aacaucagac
840acaggcuccc ugccacccaa ccuccagccu ggauauucuc cuuccccaac
ccauccuccu 900acuggacagu auacgcucuu cccucuucca cccaccuugc
ccaccccugu gguccagcuc 960cacccccugc uuccugaccc uucugcucca
acgcccaccc cuaccagccc ucuucuaaac 1020acauccuaca cccacuccca
gaaucugucu caggaagggu aa 10621504599RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
150augggacaca guaaacagau ucgaauuuua cuucugaacg aaauggagaa
acuggaaaag 60acccucuuca gacuugaaca aggguaugag cuacaguucc gauuaggccc
aacuuuacag 120ggaaaagcag uuaccgugua uacaaauuac ccauuuccug
gagaaacauu uaauagagaa 180aaauuccguu cucuggauug ggaaaaucca
acagaaagag aagaugauuc ugauaaauac 240uguaaacuua aucugcaaca
aucugguuca uuucaguauu auuuccuuca aggaaaugag 300aaaaguggug
gagguuacau aguuguggac cccauuuuac guguuggugc ugauaaucau
360gugcuacccu uggacugugu uacucuucag acauuuuuag cuaaguguuu
gggaccuuuu 420gaugaauggg aaagcagacu uaggguugca aaagaaucag
gcuacaacau gauucauuuu 480accccauugc agacucuugg acuaucuagg
ucaugcuacu cccuugccaa ucaguuagaa 540uuaaauccug acuuuucaag
accuaauaga aaguauaccu ggaaugaugu uggacagcua 600guggaaaaau
uaaaaaagga auggaauguu auuuguauua cugauguugu cuacaaucau
660acugcugcua auaguaaaug gauccaggaa cauccagaau gugccuauaa
ucuugugaau 720ucuccacacu uaaaaccugc cugggucuua gacagagcac
uuuggcguuu cuccugugau 780guugcagaag ggaaauacaa agaaaaggga
auaccugcuu ugauugaaaa ugaucaccau 840augaauucca uccgaaaaau
aauuugggag gauauuuuuc caaagcuuaa acucugggaa 900uuuuuccaag
uagaugucaa caaagcgguu gagcaauuua gaagacuucu uacacaagaa
960aauaggcgag uaaccaaguc ugauccaaac caacaccuua cgauuauuca
agauccugaa 1020uacagacggu uuggcuguac uguagauaug aacauugcac
uaacgacuuu cauaccacau 1080gacaaggggc cagcagcaau ugaagaaugc
uguaauuggu uucauaaaag aauggaggaa 1140uuaaauucag agaagcaucg
acucauuaac uaucaucagg aacaggcagu uaauugccuu 1200uugggaaaug
uguuuuauga acgacuggcu ggccaugguc caaaacuagg accugucacu
1260agaaagcauc cuuuaguuac cagguauuuu acuuucccau uugaagagau
agacuucucc 1320auggaagaau cuaugauuca ucugccaaau aaagcuuguu
uucugauggc acacaaugga 1380uggguaaugg gagaugaucc ucuucgaaac
uuugcugaac cggguucaga aguuuaccua 1440aggagagaac uuauuugcug
gggagacagu guuaaauuac gcuaugggaa uaaaccagag 1500gacuguccuu
aucucugggc acacaugaaa aaauacacug aaauaacugc aacuuauuuc
1560cagggaguac gucuugauaa cugccacuca acaccucuuc acguagcuga
guacauguug 1620gaugcugcua ggaauuugca acccaauuua uauguaguag
cugaacuguu cacaggaagu 1680gaagaucugg acaaugucuu uguuacuaga
cugggcauua guuccuuaau aagagaggca 1740augagugcau auaauaguca
ugaagagggc agauuaguuu accgauaugg aggagaaccu 1800guuggauccu
uuguucagcc cuguuugagg ccuuuaaugc cagcuauugc acaugcccug
1860uuuauggaua uuacgcauga uaaugagugu ccuauugugc auagaucagc
guaugaugcu 1920cuuccaagua cuacaauugu uucuauggca uguugugcua
guggaaguac aagaggcuau 1980gaugaauuag ugccucauca gauuucagug
guuucugaag aacgguuuua cacuaagugg 2040aauccugaag cauugccuuc
aaacacaggu gaaguuaauu uccaaagcgg cauuauugca 2100gccaggugug
cuaucaguaa acuucaucag gagcuuggag ccaaggguuu uauucaggug
2160uauguggauc aaguugauga agacauagug gcaguaacaa gacacucacc
uagcauccau 2220cagucuguug uggcuguauc uagaacugcu uucaggaauc
ccaagacuuc auuuuacagc 2280aaggaagugc cucaaaugug caucccuggc
aaaauugaag aaguaguucu ugaagcuaga 2340acuauugaga gaaacacgaa
accuuauagg aaggaugaga auucaaucaa uggaacacca 2400gauaucacag
uagaaauuag agaacauauu cagcuuaaug aaaguaaaau uguuaaacaa
2460gcuggaguug ccacaaaagg gcccaaugaa uauauucaag aaauagaauu
ugaaaacuug 2520ucuccaggaa guguuauuau auucagaguu agucuugauc
cacaugcaca agucgcuguu 2580ggaauucuuc gaaaucaucu gacacaauuc
aguccucacu uuaaaucugg cagccuagcu 2640guugacaaug cagauccuau
auuaaaaauu ccuuuugcuu cucuugccuc cagauuaacu 2700uuggcugagc
uaaaucagau ccuuuaccga ugugaaucag aagaaaagga agauggugga
2760gggugcuaug acauaccaaa cuggucagcc cuuaaauaug caggucuuca
agguuuaaug 2820ucuguauugg cagaaauaag accaaagaau gacuuggggc
auccuuuuug uaauaauuug 2880agaucuggag auuggaugau ugacuauguc
aguaaccggc uuauuucacg aucaggaacu 2940auugcugaag uugguaaaug
guugcaggcu auguucuucu accugaagca gaucccacgu 3000uaccuuaucc
cauguuacuu ugaugcuaua uuaauuggug cauauaccac ucuucuggau
3060acagcaugga agcagauguc aagcuuuguu cagaaugguu caaccuuugu
gaaacaccuu 3120ucauuggguu caguucaacu guguggagua ggaaaauucc
cuucccugcc aauucuuuca 3180ccugcccuaa uggauguacc uuauagguua
aaugagauca caaaagaaaa ggagcaaugu 3240uguguuucuc uagcugcagg
cuuaccucau uuuucuucug guauuuuccg cugcugggga 3300agggauacuu
uuauugcacu uagagguaua cugcugauua cuggacgcua uguagaagcc
3360aggaauauua uuuuagcauu ugcggguacc cugaggcaug gucucauucc
uaaucuacug 3420ggugaaggaa uuuaugccag auacaauugu cgggaugcug
ugugguggug gcugcagugu 3480auccaggauu acuguaaaau gguuccaaau
ggucuagaca uucucaagug cccaguuucc 3540agaauguauc cuacagauga
uucugcuccu uugccugcug gcacacugga ucagccauug 3600uuugaaguca
uacaggaagc aaugcaaaaa cacaugcagg gcauacaguu ccgagaaagg
3660aaugcugguc cccagauaga ucgaaacaug aaggacgaag guuuuaauau
aacugcagga 3720guugaugaag aaacaggauu uguuuaugga ggaaaucguu
ucaauugugg cacauggaug 3780gauaaaaugg gagaaaguga cagagcuaga
aacagaggaa ucccagccac accaagagau 3840gggucugcug uggaaauugu
gggccugagu aaaucugcug uucgcugguu gcuggaauua 3900uccaaaaaaa
auauuuuccc uuaucaugaa gucacaguaa aaagacaugg aaaggcuaua
3960aaggucucau augaugagug gaacagaaaa auacaagaca acuuugaaaa
gcuauuucau 4020guuuccgaag acccuucaga uuuaaaugaa aagcauccaa
aucugguuca caaacguggc 4080auauacaaag auaguuaugg agcuucaagu
ccuuggugug acuaucagcu caggccuaau 4140uuuaccauag caaugguugu
ggccccugag cucuuuacua cagaaaaagc auggaaagcu 4200uuggagauug
cagaaaaaaa auugcuuggu ccccuuggca ugaaaacuuu agauccagau
4260gauaugguuu acuguggaau uuaugacaau gcauuagaca augacaacua
caaucuugcu 4320aaagguuuca auuaucacca aggaccugag uggcuguggc
cuauugggua uuuucuucgu 4380gcaaaauuau auuuuuccag auugaugggc
ccggagacua cugcaaagac uauaguuuug 4440guuaaaaaug uucuuucccg
acauuauguu caucuugaga gauccccuug gaaaggacuu 4500ccagaacuga
ccaaugagaa ugcccaguac uguccuuuca gcugugaaac acaagccugg
4560ucaauugcua cuauucuuga gacacuuuau gauuuauag
45991514599RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 151augggacaca guaaacagau ucgaauuuua
cuucugaacg aaauggagaa acuggaaaag 60acccucuuca gacuugaaca aggguaugag
cuacaguucc gauuaggccc aacuuuacag 120ggaaaagcag uuaccgugua
uacaaauuac ccauuuccug gagaaacauu uaauagagaa 180aaauuccguu
cucuggauug ggaaaaucca acagaaagag aagaugauuc ugauaaauac
240uguaaacuua aucugcaaca aucugguuca uuucaguauu auuuccuuca
aggaaaugag 300aaaaguggug gagguuacau aguuguggac cccauuuuac
guguuggugc ugauaaucau 360gugcuacccu uggacugugu uacucuucag
acauuuuuag cuaaguguuu gggaccuuuu 420gaugaauggg aaagcagacu
uaggguugca aaagaaucag gcuacaacau gauucauuuu 480accccauugc
agacucuugg acuaucuagg ucaugcuacu cccuugccaa ucaguuagaa
540uuaaauccug acuuuucaag accuaauaga aaguauaccu ggaaugaugu
uggacagcua 600guggaaaaau uaaaaaagga auggaauguu auuuguauua
cugauguugu cuacaaucau 660acugcugcua auaguaaaug gauccaggaa
cauccagaau gugccuauaa ucuugugaau 720ucuccacacu uaaaaccugc
cugggucuua gacagagcac uuuggcguuu cuccugugau 780guugcagaag
ggaaauacaa agaaaaggga auaccugcuu ugauugaaaa ugaucaccau
840augaauucca uccgaaaaau aauuugggag gauauuuuuc caaagcuuaa
acucugggaa 900uuuuuccaag uagaugucaa caaagcgguu gagcaauuua
gaagacuucu uacacaagaa 960aauaggcgag uaaccaaguc ugauccaaac
caacaccuua cgauuauuca agauccugaa 1020uacagacggu uuggcuguac
uguagauaug aacauugcac uaacgacuuu cauaccacau 1080gacaaggggc
cagcagcaau ugaagaaugc uguaauuggu uucauaaaag aauggaggaa
1140uuaaauucag agaagcaucg acucauuaac uaucaucagg aacaggcagu
uaauugccuu 1200uugggaaaug uguuuuauga acgacuggcu ggccaugguc
caaaacuagg accugucacu 1260agaaagcauc cuuuaguuac cagguauuuu
acuuucccau uugaagagau agacuucucc 1320auggaagaau cuaugauuca
ucugccaaau aaagcuuguu uucugauggc acacaaugga 1380uggguaaugg
gagaugaucc ucuucgaaac uuugcugaac cggguucaga aguuuaccua
1440aggagagaac uuauuugcug gggagacagu guuaaauuac gcuaugggaa
uaaaccagag 1500gacuguccuu aucucugggc acacaugaaa aaauacacug
aaauaacugc aacuuauuuc 1560cagggaguac gucuugauaa cugccacuca
acaccucuuc acguagcuga guacauguug 1620gaugcugcua ggaauuugca
acccaauuua uauguaguag cugaacuguu cacaggaagu 1680gaagaucugg
acaaugucuu uguuacuaga cugggcauua guuccuuaau aagagaggca
1740augagugcau auaauaguca ugaagagggc agauuaguuu accgauaugg
aggagaaccu 1800guuggauccu uuguucagcc cuguuugagg ccuuuaaugc
cagcuauugc acaugcccug 1860uuuauggaua uuacgcauga uaaugagugu
ccuauugugc auagaucagc guaugaugcu 1920cuuccaagua cuacaauugu
uucuauggca uguugugcua guggaaguac aagaggcuau 1980gaugaauuag
ugccucauca gauuucagug guuucugaag aacgguuuua cacuaagugg
2040aauccugaag cauugccuuc aaacacaggu gaaguuaauu uccaaagcgg
cauuauugca 2100gccaggugug cuaucaguaa acuucaucag gagcuuggag
ccaaggguuu uauucaggug 2160uauguggauc aaguugauga agacauagug
gcaguaacaa gacacucacc uagcauccau 2220cagucuguug uggcuguauc
uagaacugcu uucaggaauc ccaagacuuc auuuuacagc 2280aaggaagugc
cucaaaugug caucccuggc aaaauugaag aaguaguucu ugaagcuaga
2340acuauugaga gaaacacgaa accuuauagg aaggaugaga auucaaucaa
uggaacacca 2400gauaucacag uagaaauuag agaacauauu cagcuuaaug
aaaguaaaau uguuaaacaa 2460gcuggaguug ccacaaaagg gcccaaugaa
uauauucaag aaauagaauu ugaaaacuug 2520ucuccaggaa guguuauuau
auucagaguu agucuugauc cacaugcaca agucgcuguu 2580ggaauucuuc
gaaaucaucu gacacaauuc aguccucacu uuaaaucugg cagccuagcu
2640guugacaaug cagauccuau auuaaaaauu ccuuuugcuu cucuugccuc
cagauuaacu 2700uuggcugagc uaaaucagau ccuuuaccga ugugaaucag
aagaaaagga agauggugga 2760gggugcuaug acauaccaaa cuggucagcc
cuuaaauaug caggucuuca agguuuaaug 2820ucuguauugg cagaaauaag
accaaagaau gacuuggggc auccuuuuug uaauaauuug 2880agaucuggag
auuggaugau ugacuauguc aguaaccggc uuauuucacg aucaggaacu
2940auugcugaag uugguaaaug guugcaggcu auguucuucu accugaagca
gaucccacgu 3000uaccuuaucc cauguuacuu ugaugcuaua uuaauuggug
cauauaccac ucuucuggau 3060acagcaugga agcagauguc aagcuuuguu
cagaaugguu caaccuuugu gaaacaccuu 3120ucauuggguu caguucaacu
guguggagua ggaaaauucc cuucccugcc aauucuuuca 3180ccugcccuaa
uggauguacc uuauagguua aaugagauca caaaagaaaa ggagcaaugu
3240uguguuucuc uagcugcagg cuuaccucau uuuucuucug guauuuuccg
cugcugggga 3300agggauacuu uuauugcacu uagagguaua cugcugauua
cuggacgcua uguagaagcc 3360aggaauauua uuuuagcauu ugcggguacc
cugaggcaug gucucauucc uaaucuacug 3420ggugaaggaa uuuaugccag
auacaauugu cgggaugcug ugugguggug gcugcagugu 3480auccaggauu
acuguaaaau gguuccaaau ggucuagaca uucucaagug cccaguuucc
3540agaauguauc cuacagauga uucugcuccu uugccugcug gcacacugga
ucagccauug 3600uuugaaguca uacaggaagc aaugcaaaaa cacaugcagg
gcauacaguu ccgagaaagg 3660aaugcugguc cccagauaga ucgaaacaug
aaggacgaag guuuuaauau aacugcagga 3720guugaugaag aaacaggauu
uguuuaugga ggaaaucguu ucaauugugg cacauggaug 3780gauaaaaugg
gagaaaguga cagagcuaga aacagaggaa ucccagccac accaagagau
3840gggucugcug uggaaauugu gggccugagu aaaucugcug uucgcugguu
gcuggaauua 3900uccaaaaaaa auauuuuccc uuaucaugaa gucacaguaa
aaagacaugg aaaggcuaua 3960aaggucucau augaugagug gaacagaaaa
auacaagaca acuuugaaaa gcuauuucau 4020guuuccgaag acccuucaga
uuuaaaugaa aagcauccaa aucugguuca caaacguggc 4080auauacaaag
auaguuaugg agcuucaagu ccuuggugug acuaucagcu caggccuaau
4140uuuaccauag caaugguugu ggccccugag cucuuuacua cagaaaaagc
auggaaagcu 4200uuggagauug cagaaaaaaa auugcuuggu ccccuuggca
ugaaaacuuu agauccagau 4260gauaugguuu acuguggaau uuaugacaau
gcauuagaca augacaacua caaucuugcu 4320aaagguuuca auuaucacca
aggaccugag uggcuguggc cuauugggua uuuucuucgu 4380gcaaaauuau
auuuuuccag auugaugggc ccggagacua cugcaaagac uauaguuuug
4440guuaaaaaug uucuuucccg acauuauguu caucuugaga gauccccuug
gaaaggacuu 4500ccagaacuga ccaaugagaa ugcccaguac uguccuuuca
gcugugaaac acaagccugg 4560ucaauugcua cuauucuuga gacacuuuau
gauuuauag 45991522128RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 152augagggucc
ugggugggcg cugcggggcg cugcuggcgu gucuccuccu agugcuuccc 60gucucagagg
caaacuuuug uuuauauuuu agaaaugauu uuauauacaa ccgugcaugc
120auuucuguau uggucggcuu aucuggaugc aauuuuuucu auucuauaug
cuuuuuguca 180aagcaacagg cuucacaagu ccugguuagg aagcgucgug
caaauucuuu acuugaagaa 240accaaacagg guaaucuuga aagagaaugc
aucgaagaac ugugcaauaa agaagaagcc 300agggaggucu uugaaaauga
cccggaaacg gauuauuuuu auccaaaaua cuuaguuugu 360cuucgcucuu
uucaaacugg guuauucacu gcugcacguc agucaacuaa ugcuuauccu
420gaccuaagaa gcugugucaa ugccauucca gaccagugua guccucugcc
augcaaugaa 480gauggauaua ugagcugcaa agauggaaaa gcuucuuuua
cuugcacuug uaaaccaggu 540uggcaaggag aaaaguguga auuugacaua
aaugaaugca aagaucccuc aaauauaaau 600ggagguugca gucaaauuug
ugauaauaca ccuggaaguu accacuguuc cuguaaaaau 660gguuuuguua
ugcuuucaaa uaagaaagau uguaaagaug uggaugaaug cucuuugaag
720ccaagcauuu guggcacagc ugugugcaag aacaucccag gagauuuuga
augugaaugc 780cccgaaggcu acagauauaa ucucaaauca aagucuugug
aagauauaga ugaaugcucu 840gagaacaugu gugcucagcu uugugucaau
uacccuggag guuacacuug cuauugugau 900gggaagaaag gauucaaacu
ugcccaagau cagaagaguu gugagguugu uucagugugc 960cuucccuuga
accuugacac aaaguaugaa uuacuuuacu uggcggagca guuugcaggg
1020guuguuuuau auuuaaaauu ucguuugcca gaaaucagca gauuuucagc
agaauuugau 1080uuccggacau augauucaga aggcgugaua cuguacgcag
aaucuaucga ucacucagcg 1140uggcuccuga uugcacuucg ugguggaaag
auugaaguuc agcuuaagaa ugaacauaca 1200uccaaaauca caacuggagg
ugauguuauu aauaaugguc uauggaauau ggugucugug 1260gaagaauuag
aacauaguau uagcauuaaa auagcuaaag aagcugugau ggauauaaau
1320aaaccuggac cccuuuuuaa gccggaaaau ggauugcugg aaaccaaagu
auacuuugca 1380ggauucccuc ggaaagugga aagugaacuc auuaaaccga
uuaacccucg ucuagaugga 1440uguauacgaa gcuggaauuu gaugaagcaa
ggagcuucug gaauaaagga aauuauucaa 1500gaaaaacaaa auaagcauug
ccugguuacu guggagaagg gcuccuacua uccugguucu 1560ggaauugcuc
aauuucacau agauuauaau aauguaucca gugcugaggg uuggcaugua
1620aaugugaccu ugaauauucg uccauccacg ggcacuggug uuaugcuugc
cuugguuucu 1680gguaacaaca cagugcccuu ugcugugucc uugguggacu
ccaccucuga aaaaucacag 1740gauauucugu uaucuguuga aaauacugua
auauaucgga uacaggcccu aagucuaugu 1800uccgaucaac aaucucaucu
ggaauuuaga gucaacagaa acaaucugga guugucgaca 1860ccacuuaaaa
uagaaaccau cucccaugaa gaccuucaaa gacaacuugc cgucuuggac
1920aaagcaauga aagcaaaagu ggccacauac cuggguggcc uuccagaugu
uccauucagu 1980gccacaccag ugaaugccuu uuauaauggc ugcauggaag
ugaauauuaa ugguguacag 2040uuggaucugg augaagccau uucuaaacau
aaugauauua gagcucacuc auguccauca 2100guuuggaaaa agacaaagaa uucuuuaa
21281532127RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 153augagggucc ugggugggcg cugcggggcg
cugcuggcgu gucuccuccu agugcuuccc 60gucucagagg caaacuuuug uuuauauuuu
agaaaugauu uuauauacaa ccgugcaugc 120auuucuguau uggucggcuu
aucuggaugc aauuuuuucu auucuauaug cuuuuuguca 180aagcaacagg
cuucacaagu ccugguuagg aagcgucgug caaauucuuu acuugaagaa
240accaaacagg guaaucuuga aagagaaugc aucgaagaac ugugcaauaa
agaagaagcc 300agggaggucu uugaaaauga cccggaaacg gauuauuuuu
auccaaaaua cuuaguuugu 360cuucgcucuu uucaaacugg guuauucacu
gcugcacguc agucaacuaa ugcuuauccu 420gaccuaagaa gcugugucaa
ugccauucca gaccagugua guccucugcc augcaaugaa 480gauggauaua
ugagcugcaa agauggaaaa gcuucuuuua cuugcacuug uaaaccaggu
540uggcaaggag aaaaguguga auuugacaua aaugaaugca aagaucccuc
aaauauaaau 600ggagguugca gucaaauuug ugauaauaca ccuggaaguu
accacuguuc cuguaaaaau 660gguuuuguua ugcuuucaaa uaagaaagau
uguaaagaug uggaugaaug cucuuugaag 720ccaagcauuu guggcacagc
ugugugcaag aacaucccag gagauuuuga augugaaugc 780cccgaaggcu
acagauauaa ucucaaauca aagucuugug aagauauaga ugaaugcucu
840gagaacaugu gugcucagcu uugugucaau uacccuggag guuacacuug
cuauugugau 900gggaagaaag gauucaaacu ugcccaagau cagaagaguu
gugagguugu uucagugugc 960cuucccuuga accuugacac aaaguaugaa
uuacuuuacu uggcggagca guuugcaggg 1020guuguuuuau auuuaaaauu
ucguuugcca gaaaucagca gauuuucagc agaauuugau 1080uuccggacau
augauucaga aggcgugaua cuguacgcag aaucuaucga ucacucagcg
1140uggcuccuga uugcacuucg ugguggaaag auugaaguuc agcuuaagaa
ugaacauaca 1200uccaaaauca caacuggagg ugauguuauu aauaaugguc
uauggaauau ggugucugug 1260gaagaauuag aacauaguau uagcauuaaa
auagcuaaag aagcugugau ggauauaaau 1320aaaccuggac cccuuuuuaa
gccggaaaau ggauugcugg aaaccaaagu auacuuugca 1380ggauucccuc
ggaaagugga aagugaacuc auuaaaccga uuaacccucg ucuagaugga
1440uguauacgaa gcuggaauuu gaugaagcaa ggagcuucug gaauaaagga
aauuauucaa 1500gaaaaacaaa auaagcauug ccugguuacu guggagaagg
gcuccuacua uccugguucu 1560ggaauugcuc aauuucacau agauuauaau
aauguaucca gugcugaggg uuggcaugua 1620aaugugaccu ugaauauucg
uccauccacg ggcacuggug uuaugcuugc cuugguuucu 1680gguaacaaca
cagugcccuu ugcugugucc uugguggacu ccaccucuga aaaaucacag
1740gauauucugu uaucuguuga aaauacugua auauaucgga uacaggcccu
aagucuaugu 1800uccgaucaac aaucucaucu ggaauuuaga gucaacagaa
acaaucugga guugucgaca 1860ccacuuaaaa uagaaaccau cucccaugaa
gaccuucaaa gacaacuugc cgucuuggac 1920aaagcaauga aagcaaaagu
ggccacauac cuggguggcc uuccagaugu uccauucagu 1980gccacaccag
ugaaugccuu uuauaauggc ugcauggaag ugaauauuaa ugguguacag
2040uuggaucugg augaagccau uucuaaacau aaugauauua gagcucacuc
auguccauca 2100guuuggaaaa agacaaagaa uucuuaa
21271541725RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 154augucgaucc aggagaacau aucaucccug
cagcuucggu caugggucuc uaagucccaa 60agagacuuag caaaguccau ccugauuggg
gcuccaggag ggccagcggg guaucugcgg 120cgggccagug uggcccaacu
gacccaggag cugggcacug ccuucuucca gcagcagcag 180cugccagcug
cuauggcaga caccuuccug gaacaccucu gccuacugga cauugacucc
240gagcccgugg cugcucgcag uaccagcauc auugccacca ucgggccagc
aucucgcucc 300guggagcgcc ucaaggagau gaucaaggcc gggaugaaca
uugcgcgacu caacuucucc 360cacggcuccc acgaguacca ugcugagucc
aucgccaacg uccgggaggc gguggagagc 420uuugcagguu ccccacucag
cuaccggccc guggccaucg cccuggacac caagggaccg 480gagauccgca
cugggauccu gcaggggggu ccagagucgg aaguggagcu ggugaagggc
540ucccaggugc uggugacugu ggaccccgcg uuccggacgc gggggaacgc
gaacaccgug 600uggguggacu accccaauau uguccggguc gugccggugg
ggggccgcau cuacauugac 660gacgggcuca ucucccuagu gguccagaaa
aucggcccag agggacuggu gacccaagug 720gagaacggcg gcguccuggg
cagccggaag ggcgugaacu ugccaggggc ccagguggac 780uugcccgggc
uguccgagca ggacguccga gaccugcgcu ucggggugga gcauggggug
840gacaucgucu uugccuccuu ugugcggaaa gccagcgacg uggcugccgu
cagggcugcu 900cuggguccgg aaggacacgg caucaagauc aucagcaaaa
uugagaacca cgaaggcgug 960aagagguuug augaaauccu ggaggugagc
gacggcauca ugguggcacg gggggaccua 1020ggcaucgaga ucccagcaga
gaagguuuuc cuggcucaga agaugaugau ugggcgcugc 1080aacuuggcgg
gcaagccugu ugucugugcc acacagaugc uggagagcau gauuaccaag
1140ccccggccaa cgagggcaga gacaagcgau gucgccaaug cugugcugga
uggggcugac 1200ugcaucaugc ugucagggga gacugccaag ggcaacuucc
cuguggaagc ggugaagaug 1260cagcaugcga uugcccggga ggcagaggcc
gcaguguacc accggcagcu guuugaggag 1320cuacgucggg cagcgccacu
aagccgugau cccacugagg ucaccgccau uggugcugug 1380gaggcugccu
ucaagugcug ugcugcugcc aucauugugc ugaccacaac uggccgcuca
1440gcccagcuuc ugucucggua ccgaccucgg gcagcaguca uugcugucac
ccgcucugcc 1500caggcugccc gccaggucca cuuaugccga ggagucuucc
ccuugcuuua ccgugaaccu 1560ccagaagcca ucugggcaga ugauguagau
cgccgggugc aauuuggcau ugaaagugga 1620aagcuccgug gcuuccuccg
uguuggagac cuggugauug uggugacagg cuggcgaccu 1680ggcuccggcu
acaccaacau caugcgggug cuaagcauau ccuga 17251551725RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
155augucgaucc aggagaacau aucaucccug cagcuucggu caugggucuc
uaagucccaa 60agagacuuag caaaguccau ccugauuggg gcuccaggag ggccagcggg
guaucugcgg 120cgggccagug uggcccaacu gacccaggag cugggcacug
ccuucuucca gcagcagcag 180cugccagcug cuauggcaga caccuuccug
gaacaccucu gccuacugga cauugacucc 240gagcccgugg cugcucgcag
uaccagcauc
auugccacca ucgggccagc aucucgcucc 300guggagcgcc ucaaggagau
gaucaaggcc gggaugaaca uugcgcgacu caacuucucc 360cacggcuccc
acgaguacca ugcugagucc aucgccaacg uccgggaggc gguggagagc
420uuugcagguu ccccacucag cuaccggccc guggccaucg cccuggacac
caagggaccg 480gagauccgca cugggauccu gcaggggggu ccagagucgg
aaguggagcu ggugaagggc 540ucccaggugc uggugacugu ggaccccgcg
uuccggacgc gggggaacgc gaacaccgug 600uggguggacu accccaauau
uguccggguc gugccggugg ggggccgcau cuacauugac 660gacgggcuca
ucucccuagu gguccagaaa aucggcccag agggacuggu gacccaagug
720gagaacggcg gcguccuggg cagccggaag ggcgugaacu ugccaggggc
ccagguggac 780uugcccgggc uguccgagca ggacguccga gaccugcgcu
ucggggugga gcauggggug 840gacaucgucu uugccuccuu ugugcggaaa
gccagcgacg uggcugccgu cagggcugcu 900cuggguccgg aaggacacgg
caucaagauc aucagcaaaa uugagaacca cgaaggcgug 960aagagguuug
augaaauccu ggaggugagc gacggcauca ugguggcacg gggggaccua
1020ggcaucgaga ucccagcaga gaagguuuuc cuggcucaga agaugaugau
ugggcgcugc 1080aacuuggcgg gcaagccugu ugucugugcc acacagaugc
uggagagcau gauuaccaag 1140ccccggccaa cgagggcaga gacaagcgau
gucgccaaug cugugcugga uggggcugac 1200ugcaucaugc ugucagggga
gacugccaag ggcaacuucc cuguggaagc ggugaagaug 1260cagcaugcga
uugcccggga ggcagaggcc gcaguguacc accggcagcu guuugaggag
1320cuacgucggg cagcgccacu aagccgugau cccacugagg ucaccgccau
uggugcugug 1380gaggcugccu ucaagugcug ugcugcugcc aucauugugc
ugaccacaac uggccgcuca 1440gcccagcuuc ugucucggua ccgaccucgg
gcagcaguca uugcugucac ccgcucugcc 1500caggcugccc gccaggucca
cuuaugccga ggagucuucc ccuugcuuua ccgugaaccu 1560ccagaagcca
ucugggcaga ugauguagau cgccgggugc aauuuggcau ugaaagugga
1620aagcuccgug gcuuccuccg uguuggagac cuggugauug uggugacagg
cuggcgaccu 1680ggcuccggcu acaccaacau caugcgggug cuaagcauau ccuga
17251561359RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 156auguccacug cgguccugga aaacccaggc
uugggcagga aacucucuga cuuuggacag 60gaaacaagcu auauugaaga caacugcaau
caaaauggug ccauaucacu gaucuucuca 120cucaaagaag aaguuggugc
auuggccaaa guauugcgcu uauuugagga gaaugaugua 180aaccugaccc
acauugaauc uagaccuucu cguuuaaaga aagaugagua ugaauuuuuc
240acccauuugg auaaacguag ccugccugcu cugacaaaca ucaucaagau
cuugaggcau 300gacauuggug ccacugucca ugagcuuuca cgagauaaga
agaaagacac agugcccugg 360uucccaagaa ccauucaaga gcuggacaga
uuugccaauc agauucucag cuauggagcg 420gaacuggaug cugaccaccc
ugguuuuaaa gauccugugu accgugcaag acggaagcag 480uuugcugaca
uugccuacaa cuaccgccau gggcagccca ucccucgagu ggaauacaug
540gaggaagaaa agaaaacaug gggcacagug uucaagacuc ugaaguccuu
guauaaaacc 600caugcuugcu augaguacaa ucacauuuuu ccacuucuug
aaaaguacug uggcuuccau 660gaagauaaca uuccccagcu ggaagacguu
ucucaauucc ugcagacuug cacugguuuc 720cgccuccgac cuguggcugg
ccugcuuucc ucucgggauu ucuugggugg ccuggccuuc 780cgagucuucc
acugcacaca guacaucaga cauggaucca agcccaugua uacccccgaa
840ccugacaucu gccaugagcu guugggacau gugcccuugu uuucagaucg
cagcuuugcc 900caguuuuccc aggaaauugg ccuugccucu cugggugcac
cugaugaaua cauugaaaag 960cucgccacaa uuuacugguu uacuguggag
uuugggcucu gcaaacaagg agacuccaua 1020aaggcauaug gugcugggcu
ccugucaucc uuuggugaau uacaguacug cuuaucagag 1080aagccaaagc
uucucccccu ggagcuggag aagacagcca uccaaaauua cacugucacg
1140gaguuccagc cccuguauua cguggcagag aguuuuaaug augccaagga
gaaaguaagg 1200aacuuugcug ccacaauacc ucggcccuuc ucaguucgcu
acgacccaua cacccaaagg 1260auugaggucu uggacaauac ccagcagcuu
aagauuuugg cugauuccau uaacagugaa 1320auuggaaucc uuugcagugc
ccuccagaaa auaaaguaa 13591571360RNAArtificial SequenceDescription
of Artificial Sequence Synthetic polynucleotide 157auguccacug
cgguccugga aaacccaggc uugggcagga aacucucuga cuuuggacag 60gaaacaagcu
auauugaaga caacugcaau caaaauggug ccauaucacu gaucuucuca
120cucaaagaag aaguuggugc auuggccaaa guauugcgcu uauuugagga
gaaugaugua 180aaccugaccc acauugaauc uagaccuucu cguuuaaaga
aagaugagua ugaauuuuuc 240acccauuugg auaaacguag ccugccugcu
cugacaaaca ucaucaagau cuugaggcau 300gacauuggug ccacugucca
ugagcuuuca cgagauaaga agaaagacac agugcccugg 360uucccaagaa
ccauucaaga gcuggacaga uuugccaauc agauucucag cuauggagcg
420gaacuggaug cugaccaccc ugguuuuaaa gauccugugu accgugcaag
acggaagcag 480uuugcugaca uugccuacaa cuaccgccau gggcagccca
ucccucgagu ggaauacaug 540gaggaagaaa agaaaacaug gggcacagug
uucaagacuc ugaaguccuu guauaaaacc 600caugcuugcu augaguacaa
ucacauuuuu ccacuucuug aaaaguacug uggcuuccau 660gaagauaaca
uuccccagcu ggaagacguu ucucaauucc ugcagacuug cacugguuuc
720cgccuccgac cuguggcugg ccugcuuucc ucucgggauu ucuugggugg
ccuggccuuc 780cgagucuucc acugcacaca guacaucaga cauggaucca
agcccaugua uacccccgaa 840ccugacaucu gccaugagcu guugggacau
gugcccuugu uuucagaucg cagcuuugcc 900caguuuuccc aggaaauugg
ccuugccucu cugggugcac cugaugaaua cauugaaaag 960cucgccacaa
uuuacugguu uacuguggag uuugggcucu gcaaacaagg agacuccaua
1020aaggcauaug gugcugggcu ccugucaucc uuuggugaau uacaguacug
cuuaucagag 1080aagccaaagc uucucccccu ggagcuggag aagacagcca
uccaaaauua cacugucacg 1140gaguuccagc cccuguauua cguggcagag
aguuuuaaug augccaagga ggaaaguaag 1200gaacuuugcu gccacaauac
cucggcccuu cucaguucgc uacgacccau acacccaaag 1260gauugagguc
uuggacaaua cccagcagcu uaagauuuug gcugauucca uuaacaguga
1320aauuggaauc cuuugcagug cccuccagaa aauaaaguaa
13601581359RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 158auguccacug cgguccugga aaacccaggc
uugggcagga aacucucuga cuuuggacag 60gaaacaagcu auauugaaga caacugcaau
caaaauggug ccauaucacu gaucuucuca 120cucaaagaag aaguuggugc
auuggccaaa guauugcgcu uauuugagga gaaugaugua 180aaccugaccc
acauugaauc uagaccuucu cguuuaaaga aagaugagua ugaauuuuuc
240acccauuugg auaaacguag ccugccugcu cugacaaaca ucaucaagau
cuugaggcau 300gacauuggug ccacugucca ugagcuuuca cgagauaaga
agaaagacac agugcccugg 360uucccaagaa ccauucaaga gcuggacaga
uuugccaauc agauucucag cuauggagcg 420gaacuggaug cugaccaccc
ugguuuuaaa gauccugugu accgugcaag acggaagcag 480uuugcugaca
uugccuacaa cuaccgccau gggcagccca ucccucgagu ggaauacaug
540gaggaagaaa agaaaacaug gggcacagug uucaagacuc ugaaguccuu
guauaaaacc 600caugcuugcu augaguacaa ucacauuuuu ccacuucuug
aaaaguacug uggcuuccau 660gaagauaaca uuccccagcu ggaagacguu
ucucaauucc ugcagacuug cacugguuuc 720cgccuccgac cuguggcugg
ccugcuuucc ucucgggauu ucuugggugg ccuggccuuc 780cgagucuucc
acugcacaca guacaucaga cauggaucca agcccaugua uacccccgaa
840ccugacaucu gccaugagcu guugggacau gugcccuugu uuucagaucg
cagcuuugcc 900caguuuuccc aggaaauugg ccuugccucu cugggugcac
cugaugaaua cauugaaaag 960cucgccacaa uuuacugguu uacuguggag
uuugggcucu gcaaacaagg agacuccaua 1020aaggcauaug gugcugggcu
ccugucaucc uuuggugaau uacaguacug cuuaucagag 1080aagccaaagc
uucucccccu ggagcuggag aagacagcca uccaaaauua cacugucacg
1140gaguuccagc cccuguauua cguggcagag aguuuuaaug augccaagga
gaaaguaagg 1200aacuuugcug ccacaauacc ucggcccuuc ucaguucgcu
acgacccaua cacccaaagg 1260auugaggucu uggacaauac ccagcagcuu
aagauuuugg cugauuccau uaacagugaa 1320auuggaaucc uuugcagugc
ccuccagaaa auaaaguaa 1359159132RNATobacco etch virus 159ucaacacaac
auauacaaaa acaaacgaau cucaagcaau caagcauucu acuucuauug 60cagcaauuua
aaucauuucu uuuaaagcaa aagcaauuuu cugaaaauuu ucaccauuua
120cgaacgauag cc 132160132RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 160ucaacacaac
auauacaaaa acaaacgaau cucaagcaau caagcauucu acuucuauug 60cagcaauuua
aaucauuucu uuuaaagcaa aagcaauuuu cugaaaauuu ucaccauuua
120cgaacgauag cc 132161132RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 161ucaacacaac
auauacaaaa caaacgaauc ucaagcaauc aagcauucua cuucuauugc 60agcaauuuaa
aucauuucuu uuaaagcaaa agcaauuuuc ugaaaauuuu caccauuuac
120gaacgauagc cc 132162131RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 162ucaacacaac
auauacaaaa caaacgaauc ucaagcaauc aagcauucua cuucuauugc 60agcaauuuaa
aucauuucuu uuaaagcaaa agcaauuuuc ugaaaauuuu caccauuuac
120gaacgauagc c 131163131RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 163ucaacacaac
auauacaaaa caaacgaauc ucaagcaauc aagcauucua cuucuauugc 60agcaauuuaa
aucauuucuu uuaaagcaaa agcaauuuuc ugaaaauuuu caccauuuac
120gaacgauagc c 131164131RNAArtificial SequenceDescription of
Artificial Sequence Synthetic polynucleotide 164ucaacacaac
auauacaaaa caaacgaauc ucaagcaauc aagcauucua cuucuauugc 60agcaauuuaa
aucauuucuu uuaaagcaaa agcaauuuuc ugaaaauuuu caccauuuac
120gaacgauagc c 13116529DNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 165cttcctactc
aggctttatt caaagacca 2916630RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 166aaaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 30
* * * * *