mRNA encoding engineered CFTR

Abstract

The present invention provides, among other things, improved methods and pharmaceutical compositions for treating cystic fibrosis based on codon optimized mRNA encoding an engineered or mutant Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein.

Description

MRNA ENCODING ENGINEERED CFTR CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of, and priority to U.S. Provisional Patent Application Serial No. 62 / 903,047 filed on September 20, 2019, U.S. Provisional Patent Application Serial No. 62 / 984,632, filed on March 3, 2020, and U.S. Provisional Patent Application Serial No. 63 / 021,263, filed on May 7, 2020, the contents of each of which are incorporated herein in its entirety. INCOPORATION-BY-REFERENCE OF SEQUENCE LISTING

[0002] The contents of the text file named “MRT-2115WO_SL.txt”, which was created on September 17, 2020 and is 261 KB in size, are hereby incorporated by reference in its entirety. BACKGROUND 0003) Cystic fibrosis is an autosomal inherited disorder resulting from mutation of the CFTR gene, which encodes a chloride ion channel believed to be involved in regulation of multiple other ion channels and transport systems in epithelial cells. Loss of function of CFTR results in chronic lung disease, aberrant mucus production, and dramatically reduced life expectancy. See generally Rowe et al., New Engl. J. Med. 352, 1992-2001 (2005).

[0004] Currently there is no cure for cystic fibrosis. The literature has documented numerous difficulties encountered in attempting to induce expression of CFTR in the lung. For example, viral vectors comprising CFTR DNA triggered immune responses and CF symptoms persisted after administration. Conese et al., J. Cyst. Fibros. 10 Suppl 2, S114-28 (2011); Rosenecker et al., Curr. Opin. Mol. Ther. 8, 439-45 (2006). Non-viral delivery of DNA, including CFTR DNA, has also been reported to trigger immune responses. Alton et al, Lancet 353, 947-54 (1999); Rosenecker et al., J Gene Med. 5, 49-60 (2003). Furthermore, non-viral DNA vectors encounter the additional problem that the machinery of the nuclear pore complex does not ordinarily import DNA into the nucleus, where transcription would occur. Pearson, Nature 460, 164-69 (2009). SUMMARY OF THE INVENTION

[0005] The present invention provides, among other things, pharmaceutical compositions comprising messenger RNA (mRNA) encoding an engineered or mutant Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein and methods of making and using thereof. Notably, engineered or mutant CFTR proteins described herein contain mutations that enhance the activity or stability of the protein. These pharmaceutical compositions can be used for improved treatment of cystic fibrosis.

[0006] In one aspect, the present invention provides a pharmaceutical composition for treating cystic fibrosis, comprising an mRNA encoding a Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), wherein the CFTR comprises one or more mutations that produce an activated CFTR protein.

[0007] In one aspect, a pharmaceutical composition is provided herein for treating cystic fibrosis, comprising an mRNA encoding a Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), wherein the CFTR comprises one or more mutations that produce a CFTR protein that is more stable than a wild type CFTR protein.

[0008] For example, in some embodiments, the mutant CFTR protein has a greater half- life in comparison to wild type CFTR. In some embodiments, the mutant CFTR protein has about the same half-life as a wild type CFTR. In some embodiments, the mutant CFTR protein has about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, or 24 hours greater half-life in comparison to wild type CFTR. In some embodiments, the mutant CFTR protein has more than 24 hours half-life in comparison to wild type CFTR.

[0009] Tn some embodiments, one or more mutations are located within the CFTR regulatory domain, a membrane-spanning domain (MSD), and / or a nucleotide-binding domain (NBD). Tn some embodiments, one or more mutations are located within the CFTR regulatory domain amino acid residues 634-835. In some embodiments, mutations are located within different domains.

[0010] In some embodiments, the engineered CFTR protein comprising one or more mutations results in increased activity in comparison to wild type CFTR protein.

[0011] In some embodiments, the engineered CFTR protein comprising one or more mutations results in increased trafficking in comparison to wild type CFTR protein.

[0012] In some embodiments, the engineered CFTR protein comprising one or more mutations results in increased conductivity in comparison to wild type CFTR protein. In some embodiments, the engineered CFTR protein comprising one or more mutations results in increased conductivity in the presence of modulators in comparison to wild type CFTR protein. For example, the engineered CFTR protein has increased conductivity in the presence of a potentiator, corrector and / or activator in comparison to conductivity in a wild type CFTR protein

[0013] In some embodiments, the engineered CFTR protein comprising one or more mutations has improved pharmacokinetic properties in comparison to wild type CFTR protein

[0014] In some embodiments, the engineered CFTR protein comprising one or more mutations produces an activated CFTR protein that is more stable than a wild type CFTR protein.

[0015] In some embodiments, the engineered CFTR protein comprising one or more mutations produces a CFTR protein that is more therapeutically effective for the treatment of cystic fibrosis in comparison to a wild type CFTR protein. For example, the engineered CFTR. protein has one or more of improved activity, trafficking, synergistic conductivity in the presence of modulators (e.g., potentiators, correctors and / or activators), pharmacokinetic properties, and stability in comparison to wild type CFTR protein.

[0016] In some embodiments, the CFTR comprises at least 2 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 3 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 4 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 5 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 6 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 7 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 10 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 11 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 12 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 13 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 14 mutations in amino acid residues in comparison to wild type CFTR protein. Tn some embodiments, the CFTR comprises at least 15 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 16 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 17 mutations in amino acid residues in comparison to wild type CFTR protein, Tn some embodiments, the CFTR. comprises at least 18 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 19 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises at least 20 mutations in amino acid residues in comparison to wild type CFTR protein.

[0017] In some embodiments, the CFTR comprises between 1 and 20 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises between 2 and 15 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises between 3 and 12 mutations in amino acid residues in comparison to wild type CFTR protein. In some embodiments, the CFTR comprises between 5 and 10 mutations in amino acid residues in comparison to wild type CFTR protein.

[0018] In some embodiments, the mutation is an amino acid substitution. In some embodiments, the mutation is a deletion of one or more amino acids. In some embodiments, the mutation is an insertion of one or more amino acids. In some embodiments, the mutation is an amino acid inversion. In some embodiments, the mutation is a combination of amino acid substitution, a deletion of one or more amino acids, an insertion of one or more amino acids, an amino acid inversion.

[0019] In some embodiments, one or more mutations are made at residues K14, K68, S422, S660, S670, S686, T690, $700, K710, S712, K716, S737, S753, S768, T787, T788, S790, $795, S813, K978, K1041, K1080, K1218, E1371, K190, G550, R553, R555, K1250, K464, ‘W401, F409, F430, and combinations thereof. In some embodiments, a mutation is made at residue K14. In some embodiments, a mutation is made at residue K68. In some embodiments, a mutation is made at residue S422. In some embodiments, a mutation is made at residue S660. In some embodiments, a mutation is made at residue S670. In some embodiments, a mutation is made at residue S686. In some embodiments, a mutation is made at residue S700. In some embodiments, a mutation is made at residue K710. In some embodiments, a mutation is made at residue $712. In some embodiments, a mutation is made at residue K716, In some embodiments, a mutation is made at residue S737. In some embodiments, a mutation is made at residue S753. In some embodiments, a mutation is made at residue S768. In some embodiments, a mutation is made at residue T787. In some embodiments, a mutation is made at residue T788. In some embodiments, a mutation is made at residue $790. Tn some embodiments, a mutation is made at residue $795. In some embodiments, a mutation is made at residue $813. Tn some embodiments, a mutation is made at residue K978. In some embodiments, a mutation is made at residue K1041 In some embodiments, a mutation is made at residue K1080. In some embodiments, a mutation is made at residue K1218. In some embodiments, a mutation is made at residue E1371. In some embodiments, a mutation is made at residue K190. In some embodiments, a mutation is made at residue G550. In some embodiments, a mutation is made at residue R553. In some embodiments, a mutation is made at residue R555. In some embodiments, a mutation is made at residue K1250. In some embodiments, a mutation is made at residue K464. In some embodiments, a mutation is made at residue W401. In some embodiments, a mutation is made at residue F409. In some embodiments, a mutation is made at residue F430. Accordingly, in some embodiments, one or more CFTR lysine residues are mutated. In some embodiments, one or more CFTR serine residues are mutated. In some embodiments, one or more CFTR glycine residues are mutated. In some embodiments, one or more CFTR threonine residues are mutated. In some embodiments, one or more CFTR arginine residues are mutated. In some embodiments, one or more CFTR tryptophan residues are mutated. In some embodiments, one or more CFTR threonine residues are mutated. In some embodiments, one or more CFTR glutamic acid residues are mutated. In some embodiments, one or more CFTR phenylalanine residues are mutated.

[0020] In some embodiments, CFTR comprises one or more ATP gating cycle mutations. Tn some embodiments, CFTR comprises mutations at residues W401 and E1371. In some embodiments, CFTR comprises mutations at residues K464 and E1371. In some embodiments, CFTR comprises mutations at W401, F409, and E1371. In some embodiments, CFTR comprises mutations at W401, F409, F430 and E1371. In some embodiments, CFTR comprises a mutation at K978. In some embodiments, CFTR comprises a mutation at E1371. In some embodiments, CFTR comprises mutations at residues W401 and E1371. In some embodiments, CFTR comprises mutations at W401, F409, and E1371. In some embodiments, CFTR comprises mutations at W401, F409, F430, and E1371. In some embodiments, CFTR comprises mutations at K464 and E1371.

[0021] Tn some embodiments, the one or more mutations result in a substitution to an alanine residue. In some embodiments, the one or more mutations result in a substitution to an arginine residue. In some embodiments, the one or more mutations result in a substitution to an asparagine residue. In some embodiments, the one or more mutations result in a substitution to an aspartate residue. In some embodiments, the one or more mutations result in a substitution to a cysteine residue. In some embodiments, the one or more mutations result in a substitution to a glutamate residue. In some embodiments, the one or more mutations result in a substitution to a glutamine residue. In some embodiments, the one or more mutations result in a substitution to a glycine residue. In some embodiments, the one or more mutations result in a substitution to a histidine residue. In some embodiments, the one or more mutations result in a substitution to an isoleucine residue. In some embodiments, the one or more mutations result in a substitution to a leucine residue. In some embodiments, the one or more mutations result in a substitution to a lysine residue. In some embodiments, the one or more mutations result in a substitution to a methionine residue. In some embodiments, the one or more mutations result in a substitution to a phenylalanine residue. In some embodiments, the one or more mutations result in a substitution to a proline residue. In some embodiments, the one or more mutations result in a substitution to a serine residue. In some embodiments, the one or more mutations result in a substitution to a threonine residue. In some embodiments, the one or more mutations result in a substitution to a tryptophan residue. In some embodiments, the one or more mutations result ina substitution to a tyrosine residue.

[0022] In some embodiments, one or more mutations are selected from K14R, K68R, K710R, K716R, K1041R, K1080R, K1218R, and combinations thereof.

[0023] In some embodiments, the CFTR comprises mutations selected from K14R, K68R, KI218R, K14R / K68R, K14R / K1218R, K68R / K1218R, and K14R / K68R / K1218R. Accordingly, in some embodiments, the one or more mutations is a K14R mutation. In some embodiments, the one or more mutations is a K68R mutation. In some embodiments, the one or more mutations is a K1218R mutation. In some embodiments, the one or more mutations is a K14R / K68R mutation. In some embodiments, the one or more mutations is a K14R / K1218R. mutation. In some embodiments, the one or more mutations is a K68R / K1218R mutation. In some embodiments, the one or more mutations is a K14R / K68R / K 12 18R mutation.

[0024] In some embodiments, the CFTR comprises mutations S660D / S737D / S795D / S813D, S660DD / S686D / S700D / S737D / S795D / S813D, K978C, or E1371Q. In some embodiments, the CFTR comprises mutations S660D / S737D / S795D / S813D. In some embodiments, the CFTR comprises mutations S660DD / S686D / S700D / S737D / S795D / S813D. In some embodiments, the one or more mutations is a K978C mutation. In some embodiments, the one or more mutations is a E1371Q mutation. Tn some embodiments, the CFTR comprises mutations $422D / 8660D / S670D / S686D / T690D / S700D / S712D / S753D / T787D / T788D / S790D / S795D / S813 D. In some embodiments, the CFTR comprises mutations $422D / 8660D / S670D / S686D / T690D / S700D / S712D / S737D / S753D / S768D / T787D / T788D / S790 D / S795D / S813D. 0025) In some embodiments, the one or more mutations made at residues $422, S660, S670, S686, T690, $700, $712, $737, 8753, $768, T787, T788, S790, S795, S813 is an amino acid substitution to glutamic acid (E). In some embodiments, the CFTR comprises mutations S422E / S660E / S670E / S686E / T690E / ST00E / S712E / S753E / T787E / T788E / S790E / S7T95E / S813E. In some embodiments, the CFTR comprises mutations S422E / S660E / S670E / S686E / T690E / STO0E / S7T12E / S737E / S753E / S7T68FE / T787F / T788E / S790E / S795F / SR13E. 0026] In some embodiments, the CFTR further comprises K14R mutation. In some embodiments, the CFTR further comprises E1371Q mutation. In some embodiments, the CFTR comprises mutations K14R / S422D / S660D / S670D / S686D / T690D / S700D / S712D / S753D / T787D / T788D / S790D / S795D / S813D. In some embodiments, the CFTR comprises mutations K14R / S422D / S660D / S670D / S686D / T690D / S700D / S712D / S737D / S753D / S768D / T787D / T788 D / S790D / S795D / S813D. In some embodiments, the CFTR comprises mutations K14R / S422E / S660E / S670E / S686E / T690E / S700E / S712E / S753E / T787E / T788E / S7T90E / S79SE / S813E. In some embodiments, the CFTR comprises mutations K14R / S422E / S660E / S670E / S686E / T690E / S700E / S712E / S737E / S7S3E / S768E / T787E / T788E / S790E / S79SE / S813E.

[0027] In some embodiments, the mRNA encoding CFTR is codon optimized. In some embodiments, the mRNA encoding CFTR is not codon optimized.

[0028] In some embodiments, the codon optimized CFTR mRNA further comprises a 5° untranslated region (UTR) sequence of SEQ ID NO: 4.

[0029] In some embodiments, the codon optimized CFTR mRNA further comprises a 3° untranslated region (UTR) sequence of SEQ ID NO: 5 of SEQ ID NO: 6.

[0030] In some embodiments, the codon optimized CFTR mRNA is encapsulated within a nanoparticle. In some embodiments, the nanoparticle is a liposome. In some embodiments, the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids. In some embodiments, the liposome comprises no more than three distinct lipid components. In some embodiments, one distinct lipid component is a sterol-based cationic lipid. In some embodiments, a sterol-based cationic lipid is imidazole cholesterol ester (ICE).

[0031] In some embodiments, the liposome has a size of less than about 200 nm. Tn some embodiments, the liposome has a size of less than about 150 nm. In some embodiments, the liposome has a size of less than about 120 nm. In some embodiments, the liposome has a size of less than about 110 nm. In some embodiments, the liposome has a size of less than about 100 nm. Tn some embodiments, the liposome has a size of less than about 80 nm. In some embodiments, the liposome has a size of less than about 60 nm. In some embodiments, the liposome has a size of less than about 50 nm.

[0032] In some embodiments, the pharmaceutical composition further comprises a CFTR potentiator. In some embodiments, the pharmaceutical composition further comprises a CFTR corrector. In some embodiments, the pharmaceutical composition further comprises a CFTR activator. In some embodiments, the pharmaceutical composition further comprises a CFTR potentiator, corrector and / or activator. Suitable CFTR potentiators, correctors and / or activators include ivacaftor (trade name Kalydeco®), lumacaftor (trade name Orkambi®), tezacaftor, vX- 659, VX-445, VX-152, VX-440, VX-371, VX-561, GLPG1837, GLPG2222, GLPG2737, GLPG2451, GLPG1837, PTI-428, PTI-801, PTI-808, and eluforsen. In some embodiments, the pharmaceutical composition further comprises ivacaftor. In some embodiments, the pharmaceutical composition further comprises lumacaftor. In some embodiments, the pharmaceutical composition further comprises tezacaftor. In some embodiments, the pharmaceutical composition further comprises ivacaftor, lumacaftor, tezacaftor, or a combination. Tn some embodiments, the pharmaceutical composition further comprises VX-659. In some embodiments, the pharmaceutical composition further comprises VX-445, In some embodiments, the pharmaceutical composition further comprises VX-152. In some embodiments, the pharmaceutical composition further comprises VX-440. In some embodiments, the pharmaceutical composition further comprises VX-371. In some embodiments, the pharmaceutical composition further comprises VX-561. In some embodiments, the pharmaceutical composition further comprises GLPG1837. In some embodiments, the pharmaceutical composition further comprises GLPG2222. In some embodiments, the pharmaceutical composition further comprises GLPG2737. In some embodiments, the pharmaceutical composition further comprises GLPG2451. In some embodiments, the pharmaceutical composition further comprises GLPG1837. In some embodiments, the pharmaceutical composition further comprises PTI-428. In some embodiments, the pharmaceutical composition further comprises PTI-801. In some embodiments, the pharmaceutical composition further comprises PTI-808. In some embodiments, the pharmaceutical composition further comprises eluforsen. In some embodiments, the pharmaceutical composition further comprises any combination of CFTR potentiators, correctors, and / or activators.

[0033] In one aspect, the invention provides, among other things, a polynucleotide comprising a sequence complementary to the sequence of the mRNA of the present invention.

[0034] In some embodiments, the polynucleotide is a linear or circular polynucleotide comprising deoxyribonucleotide residues.

[0035] In one aspect, the invention provides, among other things, a cultured cell comprising the polynucleotide of the present invention.

[0036] In one aspect, the invention provides a method of inducing CFTR expression in epithelial cells in a lung of a mammal comprising a step of contacting the epithelial cells in the lung of the mammal with a pharmaceutical composition of the present invention.

[0037] In some embodiments, the codon optimized CFTR mRNA is administered via pulmonary delivery. In some embodiments, the codon optimized CFTR mRNA is administered via intravenous delivery. In some embodiments, the codon optimized CFTR mRNA is administered via oral, rectal, vaginal, transmucosal, or intestinal administration; parenteral delivery, including intradermal, transdermal (topical), intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, and / or intranasal administration.

[0038] In some embodiments, the pulmonary delivery is nebulization. In some embodiments, the codon optimized CFTR mRNA is administered via aerosolization.

[0039] In some embodiments, treating the subject is achieved at a lower therapeutically effective dose in comparison to treating the subject with an mRNA encoding a wild type CFTR

[0040] In some embodiments, treating the subject in need results in shorter nebulization times to administer a therapeutically effective dose in comparison to treating with an mRNA encoding wild type CFTR.

[0041] In one aspect, the invention provides, among other things, a nucleic acid encoding a modified Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein, wherein the modified CFTR protein comprises one or more amino acid substitutions to glutamic acid.

[0042] Tn some embodiments, one or more amino acid substitutions occur at serine residues. In some embodiments, one or more amino acid substitutions occur at threonine residues.

[0043] In some embodiments, one or more amino acid substitutions are located within the CFTR regulatory domain. In some embodiments, one or more amino acid substitutions are located at amino acid residues 440-820. In some embodiments, one or more amino acid substitutions occur at residue $422, $660, S670, $686, T690, $700, S712, $737, S753, $768, T787, T788, S790, S795, and / or S813.

[0044] In some embodiments, a modified CFTR protein comprises one or more mutations of S422E, S660E, S670E, S686E, T690E, S700E, S712E, S737E, S753E, S768E, T787E, T788E, S790E, S795E, and / or S813E. In some embodiments, a modified CFTR protein comprises mutations S422E / S660E / S670E / S686E / T690E / ST00E / S712E / S753E / T787E / T788E / S790E / S795E / S813E. In some embodiments, a modified CFTR protein comprises mutations S422E / S660E / S670E / S686E / T690E / ST00E / S712E / S737E / S753E / S768E / T787E / T788E / S790E / S795F / SR13F.

[0045] In some embodiments, a modified CFTR protein further comprises K14R mutation. In some embodiments, a modified CFTR protein further comprises E1371Q mutation.

[0046] In one aspect, the invention provides, among other things, a nucleic acid encoding a modified Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein, wherein the modified CFTR protein comprises K14R and E1371Q mutations.

[0047] In some embodiments, a modified CFTR protein further comprises one or more mutations in the CFTR regulatory domain, a membrane-spanning domain (MSD), and / or a nucleotide-binding domain.

[0048] Tn some embodiments, a nucleic acid is DNA, cDNA, RNA, mRNA or PNA. In some embodiments, a nucleic acid is DNA. In some embodiments, a nucleic acid is cDNA. In some embodiments, a nucleic acid is RNA. In some embodiments, a nucleic acid is mRNA. In some embodiments, a nucleic acid is PNA. In some embodiments, a nucleic acid is PCNA. In some embodiments, a nucleic acid is MCNA.

[0049] In one aspect, the invention provides, among other things, an adeno-associated virus (AAV) vector comprising a nucleic acid encoding a modified Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein.

[0050] Tn some embodiments, an AAV vector is packaged in a virus or viral particle. In some embodiments, a virus particle is pseudotyped. In some embodiments, a virus is a retrovirus, adenovirus, adeno-associate virus, herpes virus, or lentivirus.

[0051] In one aspect, the invention provides, among other things, a modified CFTR protein comprising one or more amino acid substitutions to glutamic acid.

[0052] In one aspect, the invention provides, among other things, a method of treating cystic fibrosis comprising administering to a subject in need of treatment a modified CFTR protein

[0053] In one aspect, the invention provides, among other things, a method of inducing CFTR expression in a subject, the method comprising administering the AAV vector. In one aspect, the invention provides, among other things, a method of treating cystic fibrosis comprising administering to a subject in need of treatment the AAV vector. BRIEF DESCRIPTION OF THE DRAWING

[0054] The drawings are for illustration purposes only not for limitation.

[0055] FIG. 1 shows a schematic that illustrates the regulation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel activity.

[0056] FIG. 2A and FIG. 2B show schematics of exemplary engineered CFTR proteins with mutations that activate and / or stabilize and / or enhance CFTR protein expression. FIG. 2A shows a schematic model and exemplary R domain phosphomimetic mutations. FIG. 2B shows a schematic model of ATP-gating mutations. FIG. 2C shows exemplary stability / expression- enhancing mutations. FIG. 2D shows exemplary engineered CFTR stability mutant protein expression relative to WT.

[0057] FIG. 3A is a gel that shows mutated CFTR constructs are expressed to similar levels as compared with wild type CFTR in cultured HEK cells.

[0058] FIG. 3B shows an exemplary graph of short circuit current depicting chloride ion flux over time through WT CFTR and engineered CFTR mutant protein channels in an Ussing Chamber Assay at a high dose of 0.5 pg.

[0059] FIG. 4 shows an exemplary graph of short circuit current depicting chloride ion flux over time through WT CFTR and engineered CFTR mutant protein channels in an Ussing Chamber Assay at a low dose of 0.35 ug.

[0060] FIG. 5 shows exemplary results of a graph of a time course of short-circuit current, a measure of chloride ion transport through engineered CFTR protein E at doses ranging from 0.05-0.35 pg.

[0061] FIG. 6 shows exemplary results of a graph of a time course of short-circuit current, a measure of chloride ion transport through WT CFTR protein upon ecsin permeabilization of the membrane using doses from 10-25 uM ecsin in the presence and absence of ATP.

[0062] FIG. 7 shows exemplary results of a graph of a time course of short-circuit current, a measure of chloride ion transport through WT CFTR protein at a dose of 10 uM ecsin in an Ussing Chamber Assay.

[0063] FIG. 8 shows an exemplary graph of short circuit current depicting chloride ion flux over time through WT CFTR and engineered phosphomimetic CFTR mutant protein channels in an Ussing Chamber Assay.

[0064] FIG. 9A shows exemplary graph of short-circuit conductivity measured by Ussing chamber using FRT cells transfected with wild-type, 13D, and 13E CFTR mRNA. FIG. 9B shows exemplary graph of short-circuit conductivity measured by Ussing chamber using FRT cells transfected with wild-type, 15D, and 15E CFTR mRNA. Arrowheads indicate addition of forskolin (f), VX-770 Ivacaftor (v), or CFTR inhibitor 172. Error bars represent s.e.m. with N=3 or4.

[0065] FIG. 10 shows an exemplary bar graph depicting maximum activated current for various trafficking CFTR mutant proteins listed in Table 10, as determined by an Ussing Chamber assay.

[0066] FIG. 11A shows an exemplary a Western blot illustrating expression of CFTR proteins (15E, 13E, E1371Q variants, and wild-type CFTR) in HEK293 cells. FIG. 11B is a quantification of expression levels as shown in FIG. 11A. FIG. 11C shows exemplary graph of short-circuit conductivity measured by Ussing chamber using FRT cells transfected with wild- type, 15E, 15E / K14R, 13D, 13D / K14R, 13E and 13E / K14R CFTR mRNAs. FIG. 11D is a bar graph representation of maximum activated current for various CFTR constructs tested in FIG. 11C. FIG. 11E is a series of graphs that show short-circuit conductivity measured by Ussing chamber using FRT cells that were transfected with WT, 13D, 13E, 15D or 15E CFTR mRNAs as indicated.

[0067] FIG. 12A is a table of currents depicting chloride ion flux at baseline and after forskolin, VX-770, and CFTRinh-172 additions. FIG. 12B is an exemplary graph of short- circuit conductivity illustrating, among other things, that alanine substitution at phosphomimetic sites reduces forskolin-induced chloride conductance in FRT cells.

[0068] FIG. 13A shows exemplary graph of short-circuit conductivity measured by Ussing chamber using FRT cells transfected with wild-type, E1371Q and E1371Q / K14R CFTR mRNAs. Arrowheads indicate addition of forskolin (f), VX-770 Ivacaftor (v), or CFTR inhibitor 172. Error bars represent s.e.m. with N=3 or 4. FIG. 13B is a bar graph representation of activity and expression of E1371Q CFTR mutant and E1371Q / K14R CFTR mutant compared to the wild-type CFTR protein.

[0069] FIG. 14A and FIG. 14B is a bar graph representation of maximum activation current for various CFTR constructs in Ussing chamber assay at 22 and 44 hours. FIG. 14C is an exemplary bar graphs showing that removal of ubiquitination site at N terminus alters channel activity when combined with other substitutions.

[0070] FIG. 15A depicts a schematic that shows CFTR that contains mutations at the indicated amino acid residues. FIG. 15B depicts a series of graphs that shows channel relaxation times for the indicated CFTR mutants. Fig, 15C depicts a series of graphs that show measurements of short-circuit conductivity measured by Ussing chamber of CFTR variants.

[0071] FIG. 16A depicts a graph that shows short-circuit conductivity measured by Ussing chamber using FRT cells that were transfected with CFTR mRNAs that had phosphomimetic CFTR mutations. FIG. 16B is a graph that shows forskolin EC50 (nM) in CFTR mRNAs that had varying number of phosphomimetic mutations.

[0072] FIG. 17 depicts a graph that shows forskolin sensitivity of CFTR mRNAs that had specific amino acid substitutions as assessed by short-circuit conductivity measured by Ussing chamber using FRT cells that were transfected with CFTR mRNAs identified in the graph.

[0073] FIG. 18A is a gel that depicts the banding patterns of various CFTR sequences. FIG. 18B is a bar graph that depicts CFTR mRNA potency of various CFTR sequences. FIG. 18C is a bar graph that depicts the relative expression of various CFTR sequences in the C band.

[0074] FIG. 19 is a bar graph that depicts the results of an assay to assess cytotoxicity of various CFTR mRNA sequences.

[0075] FIG. 204, 20B, 20C, 20D, 20E, and 20F are a series of graphs that show short- circuit conductivity measured by Ussing chamber of various codon-optimized and non-codon optimized CFTR constructs. DEFINITIONS 0076) In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference.

[0077] Approximately or about: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0078] As used herein, the term “batch” refers to a quantity or amount of mRNA synthesized at one time, e.g., produced according to a single manufacturing order during the same cycle of manufacture. A batch may refer to an amount of mRNA synthesized in one reaction that occurs via a single aliquot of enzyme and / or a single aliquot of DNA template for continuous synthesis under one set of conditions. Tn some embodiments, a batch would include the mRNA produced from a reaction in which not all reagents and / or components are supplemented and / or replenished as the reaction progresses. The term “not in a single batch” would not mean mRNA synthesized at different times that are combined to achieve the desired amount. 10079) Delivery: As used herein, the term “delivery” encompasses both local and systemic delivery. For example, delivery of mRNA encompasses situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also referred to as “local distribution” or “local delivery”), and situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into patient’s circulation system (e.g., serum) and systematically distributed and taken up by other tissues (also referred to as “systemic distribution” or “systemic delivery). In some embodiments, delivery is pulmonary delivery, e.g., comprising nebulization.

[0080] Encapsulation: As used herein, the term “encapsulation,” or grammatical equivalent, refers to the process of confining an mRNA molecule within a nanoparticle.

[0081] Engineered or mutant: As used herein, the terms “engineered” or “ mutant”, or grammatical equivalents refer to a nucleotide or protein sequence comprising one or more modifications compared to its naturally-occurring sequence, including but not limited to deletions, insertions of heterologous nucleic acids or amino acids, inversions, substitutions, or combinations thereof.

[0082] Expression: As used herein, “expression” of a nucleic acid sequence refers to translation of an mRNA into a polypeptide, assemble multiple polypeptides (e.g., heavy chain or light chain of antibody) into an intact protein (e.g., antibody) and / or post-translational modification of a polypeptide or fully assembled protein (e.g, antibody). In this application, the terms “expression” and “production,” and grammatical equivalents, are used interchangeably.

[0083] Functional: As used herein, a “functional” biological molecule is a biological molecule in a form in which it exhibits a property and / or activity by which it is characterized.

[0084] Half-life: As used herein, the term “half-life” is the time required for a quantity such as nucleic acid or protein concentration or activity to fall to half of its value as measured at the beginning of a time period.

[0085] Improve, increase, or reduce: As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control subject (or multiple control subject) in the absence of the treatment described herein. A “control subject” is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

[0086] Impurities. As used herein, the term “impurities” refers to substances inside a confined amount of liquid, gas, or solid, which differ from the chemical composition of the target material or compound. Impurities are also referred to as contaminants.

[0087] In Vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, efc., rather than within a multi-cellular organism.

[0088] In Vivo: As used herein, the term “in vivo” refers to events that occur within a multi-cellular organism, such as a human and a non-human animal. In the context of cell-based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

[0089] Isolated: As used herein, the term “isolated” refers to a substance and / or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and / or in an experimental setting), and / or (2) produced, prepared, and / or manufactured by the hand of man. Isolated substances and / or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. Tn some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is “pure” if it is substantially free of other components. As used herein, calculation of percent purity of isolated substances and / or entities should not include excipients (e.g.. buffer, solvent, water, etc.) 10090] messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” refers to a polynucleotide that encodes at least one polypeptide. mRNA as used herein encompasses both modified and unmodified RNA. mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, mRNA can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, ec. An mRNA sequence is presented in the 5' to 3’ direction unless otherwise indicated.

[0091] Nucleic acid: As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and / or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester linkage. In some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and / or nucleosides). In some embodiments, “nucleic acid” refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, “nucleic acid” encompasses RNA as well as single and / or double-stranded DNA and / or cDNA. Furthermore, the terms “nucleic acid,” “DNA,” “RNA,” and / or similar terms include nucleic acid analogs, i.e., analogs having other than a phosphodiester backbone. For example, the so-called “peptide nucleic acids,” which are known in the art and have peptide bonds instead of phosphodiester bonds in the backbone, are considered within the scope of the present invention. The term “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and / or encode the same amino acid sequence. Nucleotide sequences that encode proteins and / or RNA may include introns. Nucleic acids can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, nucleic acids can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, etc. A nucleic acid sequence is presented in the 5' to 3 direction unless otherwise indicated. In some embodiments, a nucleic acid is or comprises natural nucleosides (e.g., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine); nucleoside analogs (e.g., 2-aminoadenosine, 2- thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2- aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)- methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases): intercalated bases; modified sugars (e.g., 2’ -fluororibose, ribose, 2’- deoxyribose, arabinose, and hexose); and / or modified phosphate groups (e.g., phosphorothioates and 5'-N-phosphoramidite linkages). In some embodiments, the present invention is specifically directed to “unmodified nucleic acids,” meaning nucleic acids (e.g., polynucleotides and residues, including nucleotides and / or nucleosides) that have not been chemically modified in order to facilitate or achieve delivery. In some embodiments, the nucleotides T and U are used interchangeably in sequence descriptions.

[0092] Patient: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. A human includes pre- and post-natal forms.

[0093] Pharmaceutically acceptable: The term “pharmaceutically acceptable” as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0094] Stable: As used herein, the term “stable” protein or its grammatical equivalents refer to protein that retains its physical stability and / or biological activity. In one embodiment, protein stability is determined based on the percentage of monomer protein in the solution, ata low percentage of degraded (e.g., fragmented) and / or aggregated protein. In one embodiment, a stable engineered protein retains or exhibits an enhanced half-life as compared to a wild-type protein. In one embodiment, a stable engineered protein is less prone to ubiquitination that leads to proteolysis as compared to a wild-type protein. 0095] Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

[0096] Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and / or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0097] Treating: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and / or reduce incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease. DETAILED DESCRIPTION

[0098] The present invention provides, among other things, improved methods and pharmaceutical compositions for treating cystic fibrosis using messenger RNA (mRNA) encoding an engineered or mutant Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein. In some embodiments, the mRNA is a codon-optimized mRNA. In particular embodiments, the engineered or mutant CFTR proteins achieve higher activity or stability than the wild-type CFTR protein. mRNAs disclosed herein encoding engineered or mutant CFTR proteins are particularly useful for treating cystic fibrosis by mRNA therapeutics. Cystic Fibrosis

[0099] The present invention may be used to treat a subject who is suffering from or susceptible to cystic fibrosis. Cystic fibrosis is a genetic disorder characterized by mutations in the gene for Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). The CFTR protein functions as a channel across the membrane of cells that produce mucus, sweat, saliva, tears, and digestive enzymes. The channel transports negatively charged particles called chloride ions into and out of cells. The transport of chloride ions helps control the movement of water in tissues, which is necessary for the production of thin, freely flowing mucus. Mucus is a slippery substance that lubricates and protects the lining of the airways, digestive system, reproductive system, and other organs and tissues.

[0100] Respiratory symptoms of cystic fibrosis include: a persistent cough that produces thick mucus (sputum), wheezing, breathlessness, exercise intolerance, repeated lung infections and inflamed nasal passages or a stuffy nose. Digestive symptoms of cystic fibrosis include: foul-smelling, greasy stools, poor weight gain and growth, intestinal blockage, particularly in newborns (meconium ileus), and severe constipation. Codon Optimized mRNA Encoding CFTR

[0101] In some embodiments, the present invention provides methods and compositions for delivering codon optimized mRNA encoding CFTR to a subject for the treatment of cystic fibrosis. A suitable codon optimized CFTR mRNA encodes any full length, fragment or portion of a CFTR protein which can be substituted for naturally-occurring CFTR protein activity and / or reduce the intensity, severity, and / or frequency of one or more symptoms associated with cystic fibrosis.

[0102] Tn some embodiments, a suitable codon optimized mRNA sequence is an mRNA sequence encoding a human CFTR (hCFTR) protein. Exemplary codon optimized CFTR mRNA coding sequence and the corresponding amino acid sequence are shown in Table 1: Table 1. Exemplary Codon-Optimized Human CFTR SEQ ID AUGCAACGCUCUCCUCUUGAAAAGGCCUCGGUGGUGUCCAAGCUCUU SEQID | cUUCUCGUGGACUAGACCCAUCCUGAGAAAGGGGUACAGACAGCGCU Noa UGGAGCUGUCCGAUAUCUAUCAAAUCCCUUCCGUGGACUCCGCGGAC AACCUGUCCGAGAAGCUCGAGAGAGAAUGGGACAGAGAACUCGCCUC AAAGAAGAACCCGAAGCUGAUUAAUGCGCUUAGGCGGUGCUUUUUC UGGCGGUUCAUGUUCUACGGCAUCUUCCUCUACCUGGGAGAGGUCAC CAAGGCCGUGCAGCCCCUGUUGCUGGGACGGAUUAUUGCCUCCUACG ACCCCGACAACAAGGAAGAAAGAAGCAUCGCUAUCUACUUGGGCAUC GGUCUGUGCCUGCUUUUCAUCGUCCGGACCCUCUUGUUGCAUCCUGC UAUUUUCGGCCUGCAUCACAUUGGCAUGCAGAUGAGAAUUGCCAUG UUUUCCCUGAUCUACAAGAAAACUCUGAAGCUCUCGAGCCGCGUGCU UGACAAGAUUUCCAUCGGCCAGCUCGUGUCCCUGCUCUCCAACAAUC UGAACAAGUUCGACGAGGGCCUCGCCCUGGCCCACUUCGUGUGGAUC GCCCCUCUGCAAGUGGCGCUUCUGAUGGGCCUGAUCUGGGAGCUGCU GCAAGCCUCGGCAUUCUGUGGGCUUGGAUUCCUGAUCGUGCUGGCAC UGUUCCAGGCCGGACUGGGGCGGAUGAUGAUGAAGUACAGGGACCA GAGAGCCGGAAAGAUUUCCGAACGGCUGGUGAUCACUUCGGAAAUG AUCGAAAACAUCCAGUCAGUGAAGGCCUACUGCUGGGAAGAGGCCAU GGAAAAGAUGAUUGAAAACCUCCGGCAAACCGAGCUGAAGCUGACCC GCAAGGCCGCUUACGUGCGCUAUUUCAACUCGUCCGCUUUCUUCUUC UCCGGGUUCUUCGUGGUGUUUCUCUCCGUGCUCCCCUACGCCCUGAU UAAGGGAAUCAUCCUCAGGAAGAUCUUCACCACCAUUUCCUUCUGUA UCGUGCUCCGCAUGGCCGUGACCCGGCAGUUCCCAUGGGCCGUGCAG ACUUGGUACGACUCCCUGGGAGCCAUUAACAAGAUCCAGGACUUCCU UCAAAAGCAGGAGUACAAGACCCUCGAGUACAACCUGACUACUACCG AGGUCGUGAUGGAAAACGUCACCGCCUUUUGGGAGGAGGGAUUUGG CGAACUGUUCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGACCU CGAACGGUGACGACUCCCUCUUCUUUUCAAACUUCAGCCUGCUCGGG ACGCCCGUGCUGAAGGACAUUAACUUCAAGAUCGAAAGAGGACAGCU CCUGGCGGUGGCCGGAUCGACCGGAGCCGGAAAGACUUCCCUGCUGA UGGUGAUCAUGGGAGAGCUUGAACCUAGCGAGGGAAAGAUCAAGCA CUCCGGCCGCAUCAGCUUCUGUAGCCAGUUUUCCUGGAUCAUGCCCG GAACCAUUAAGGAAAACAUCAUCUUCGGCGUGUCCUACGAUGAAUAC CGCUACCGGUCCGUGAUCAAAGCCUGCCAGCUGGAAGAGGAUAUUUC AAAGUUCGCGGAGAAAGAUAACAUCGUGCUGGGCGAAGGGGGUAUU ACCUUGUCGGGGGGCCAGCGGGCUAGAAUCUCGCUGGCCAGAGCCGU GUAUAAGGACGCCGACCUGUAUCUCCUGGACUCCCCCUUCGGAUACC UGGACGUCCUGACCGAAAAGGAGAUCUUCGAAUCGUGCGUGUGCAA GCUGAUGGCUAACAAGACUCGCAUCCUCGUGACCUCCAAAAUGGAGC ACCUGAAGAAGGCAGACAAGAUUCUGAUUCUGCAUGAGGGGUCCUCC UACUUUUACGGCACCUUCUCGGAGUUGCAGAACUUGCAGCCCGACUU CUCAUCGAAGCUGAUGGGUUGCGACAGCUUCGACCAGUUCUCCGCCG AAAGAAGGAACUCGAUCCUGACGGAAACCUUGCACCGCUUCUCUUUG GAAGGCGACGCCCCUGUGUCAUGGACCGAGACUAAGAAGCAGAGCUU CAAGCAGACCGGGGAAUUCGGCGAAAAGAGGAAGAACAGCAUCUUG AACCCCAUUAACUCCAUCCGCAAGUUCUCAAUCGUGCAAAAGACGCC ACUGCAGAUGAACGGCAUUGAGGAGGACUCCGACGAACCCCUUGAGA GGCGCCUGUCCCUGGUGCCGGACAGCGAGCAGGGAGAAGCCAUCCUG CCUCGGAUUUCCGUGAUCUCCACUGGUCCGACGCUCCAAGCCCGGCG GCGGCAGUCCGUGCUGAACCUGAUGACCCACAGCGUGAACCAGGGCC AAAACAUUCACCGCAAGACUACCGCAUCCACCCGGAAAGUGUCCCUG GCACCUCAAGCGAAUCUUACCGAGCUCGACAUCUACUCCCGGAGACU GUCGCAGGAAACCGGGCUCGAAAUUUCCGAAGAAAUCAACGAGGAG GAUCUGAAAGAGUGCUUCUUCGACGAUAUGGAGUCGAUACCCGCCGU GACGACUUGGAACACUUAUCUGCGGUACAUCACUGUGCACAAGUCAU UGAUCUUCGUGCUGAUUUGGUGCCUGGUGAUUUUCCUGGCCGAGGU CGCGGCCUCACUGGUGGUGCUCUGGCUGUUGGGAAACACGCCUCUGC AAGACAAGGGAAACUCCACGCACUCGAGAAACAACAGCUAUGCCGUG AUUAUCACUUCCACCUCCUCUUAUUACGUGUUCUACAUCUACGUCGG AGUGGCGGAUACCCUGCUCGCGAUGGGUUUCUUCAGAGGACUGCCGC UGGUCCACACCUUGAUCACCGUCAGCAAGAUUCUUCACCACAAGAUG UUGCAUAGCGUGCUGCAGGCCCCCAUGUCCACCCUCAACACUCUGAA GGCCGGAGGCAUUCUGAACAGAUUCUCCAAGGACAUCGCUAUCCUGG ACGAUCUCCUGCCGCUUACCAUCUUUGACUUCAUCCAGCUGCUGCUG AUCGUGAUUGGAGCAAUCGCAGUGGUGGCGGUGCUGCAGCCUUACA UUUUCGUGGCCACUGUGCCGGUCAUUGUGGCGUUCAUCAUGCUGCGG GCCUACUUCCUCCAAACCAGCCAGCAGCUGAAGCAACUGGAAUCCGA GGGACGAUCCCCCAUCUUCACUCACCUUGUGACGUCGUUGAAGGGAC UGUGGACCCUCCGGGCUUUCGGACGGCAGCCCUACUUCGAAACCCUC UUCCACAAGGCCCUGAACCUCCACACCGCCAAUUGGUUCCUGUACCU GUCCACCCUGCGGUGGUUCCAGAUGCGCAUCGAGAUGAUUUUCGUCA UCUUCUUCAUCGCGGUCACAUUCAUCAGCAUCCUGACUACCGGAGAG GGAGAGGGACGGGUCGGAAUAAUCCUGACCCUCGCCAUGAACAUUAU GAGCACCCUGCAGUGGGCAGUGAACAGCUCGAUCGACGUGGACAGCC UGAUGCGAAGCGUCAGCCGCGUGUUCAAGUUCAUCGACAUGCCUACU GAGGGAAAACCCACUAAGUCCACUAAGCCCUACAAAAAUGGCCAGCU GAGCAAGGUCAUGAUCAUCGAAAACUCCCACGUGAAGAAGGACGAU AUUUGGCCCUCCGGAGGUCAAAUGACCGUGAAGGACCUGACCGCAAA GUACACCGAGGGAGGAAACGCCAUUCUCGAAAACAUCAGCUUCUCCA UUUCGCCGGGACAGCGGGUCGGCCUUCUCGGGCGGACCGGUUCCGGG AAGUCAACUCUGCUGUCGGCUUUCCUCCGGCUGCUGAAUACCGAGGG GGAAAUCCAAAUUGACGGCGUGUCUUGGGAUUCCAUUACUCUGCAGC AGUGGCGGAAGGCCUUCGGCGUGAUCCCCCAGAAGGUGUUCAUCUUC UCGGGUACCUUCCGGAAGAACCUGGAUCCUUACGAGCAGUGGAGCGA CCAAGAAAUCUGGAAGGUCGCCGACGAGGUCGGCCUGCGCUCCGUGA UUGAACAAUUUCCUGGAAAGCUGGACUUCGUGCUCGUCGACGGGGG AUGUGUCCUGUCGCACGGACAUAAGCAGCUCAUGUGCCUCGCACGGU CCGUGCUCUCCAAGGCCAAGAUUCUGCUGCUGGACGAACCUUCGGCC SEQ ID NO:2 CACCUGGAUCCGGUCACCUACCAGAUCAUCAGGAGGACCCUGAAGCA GGCCUUUGCCGAUUGCACCGUGAUUCUCUGCGAGCACCGCAUCGAGG CCAUGCUGGAGUGCCAGCAGUUCCUGGUCAUCGAGGAGAACAAGGUC CGCCAAUACGACUCCAUUCAAAAGCUCCUCAACGAGCGGUCGCUGUU CAGACAAGCUAUUUCACCGUCCGAUAGAGUGAAGCUCUUCCCGCAUC GGAACAGCUCAAAGUGCAAAUCGAAGCCGCAGAUCGCAGCCUUGAAG GAAGAGACUGAGGAAGAGGUGCAGGACACCCGGCUUUAA AUGCAGCGGUCCCCGCUCGAAAAGGCCAGUGUCGUGUCCAAACUCUU » | CUUCUCAUGGACUCGGCCUAUCCUUAGAAAGGGGUAUCGGCAGAGGC UUGAGUUGUCUGACAUCUACCAGAUCCCCUCGGUAGAUUCGGCGGAU AACCUCUCGGAGAAGCUCGAACGGGAAUGGGACCGCGAACUCGCGUC UAAGAAAAACCCGAAGCUCAUCAACGCACUGAGAAGGUGCUUCUUCU GGCGGUUCAUGUUCUACGGUAUCUUCUUGUAUCUCGGGGAGGUCAC AAAAGCAGUCCAACCCCUGUUGUUGGGUCGCAUUAUCGCCUCGUACG ACCCCGAUAACAAAGAAGAACGGAGCAUCGCGAUCUACCUCGGGAUC GGACUGUGUUUGCUUUUCAUCGUCAGAACACUUUUGUUGCAUCCAGC AAUCUUCGGCCUCCAUCACAUCGGUAUGCAGAUGCGAAUCGCUAUGU UUAGCUUGAUCUACAAAAAGACACUGAAACUCUCGUCGCGGGUGUU GGAUAAGAUUUCCAUCGGUCAGUUGGUGUCCCUGCUUAGUAAUAAC | CUCAACAAAUUCGAUGAGGGACUGGCGCUGGCACAUUUCGUGUGGA UUGCCCCGUUGCAAGUCGCCCUUUUGAUGGGCCUUAUUUGGGAGCUG UUGCAGGCAUCUGCCUUUUGUGGCCUGGGAUUUCUGAUUGUGUUGG CAUUGUUUCAGGCUGGGCUUGGGCGGAUGAUGAUGAAGUAUCGCGA CCAGAGAGCGGGUAAAAUCUCGGAAAGACUCGUCAUCACUUCGGAAA UGAUCGAAAACAUCCAGUCGGUCAAAGCCUAUUGCUGGGAAGAAGC UAUGGAGAAGAUGAUUGAAAACCUCCGCCAAACUGAGCUGAAACUG ACCCGCAAGGCGGCGUAUGUCCGGUAUUUCAAUUCGUCAGCGUUCUU CUUUUCCGGGUUCUUCGUUGUCUUUCUCUCGGUUUUGCCUUAUGCCU UGAUUAAGGGGAUUAUCCUCCGCAAGAUUUUCACCACGAUUUCGUUC UGCAUUGUAUUGCGCAUGGCAGUGACACGGCAAUUUCCGUGGGCCGU GCAGACAUGGUAUGACUCGCUUGGAGCGAUCAACAAAAUCCAAGACU UCUUGCAAAAGCAAGAGUACAAGACCCUGGAGUACAAUCUUACUACU ACGGAGGUAGUAAUGGAGAAUGUGACGGCUUUUUGGGAAGAGGGUU UUGGAGAACUGUUUGAGAAAGCAAAGCAGAAUAACAACAACCGCAA GACCUCAAAUGGGGACGAUUCCCUGUUUUUCUCGAACUUCUCCCUGC UCGGAACACCCGUGUUGAAGGACAUCAAUUUCAAGAUUGAGAGGGG ACAGCUUCUCGCGGUAGCGGGAAGCACUGGUGCGGGAAAAACUAGCC UCUUGAUGGUGAUUAUGGGGGAGCUUGAGCCCAGCGAGGGGAAGAU UAAACACUCCGGGCGUAUCUCAUUCUGUAGCCAGUUUUCAUGGAUCA UGCCCGGAACCAUUAAAGAGAACAUCAUUUUCGGAGUAUCCUAUGA UGAGUACCGAUACAGAUCGGUCAUUAAGGCGUGCCAGUUGGAAGAG GACAUUUCUAAGUUCGCCGAGAAGGAUAACAUCGUCUUGGGAGAAG GGGGUAUUACAUUGUCGGGAGGGCAGCGAGCGCGGAUCAGCCUCGCG AGAGCGGUAUACAAAGAUGCAGAUUUGUAUCUGCUUGAUUCACCGU UUGGAUACCUCGACGUAUUGACAGAAAAAGAAAUCUUCGAGUCGUG CGUGUGUAAACUUAUGGCUAAUAAGACGAGAAUCCUGGUGACAUCA AAAAUGGAACACCUUAAGAAGGCGGACAAGAUCCUGAUCCUCCACGA AGGAUCGUCCUACUUUUACGGCACUUUCUCAGAGUUGCAAAACUUGC AGCCGGACUUCUCAAGCAAACUCAUGGGGUGUGACUCAUUCGACCAG UUCAGCGCGGAACGGCGGAACUCGAUCUUGACGGAAACGCUGCACCG AUUCUCGCUUGAGGGUGAUGCCCCGGUAUCGUGGACCGAGACAAAGA AGCAGUCGUUUAAGCAGACAGGAGAAUUUGGUGAGAAAAGAAAGAA CAGUAUCUUGAAUCCUAUUAACUCAAUUCGCAAGUUCUCAAUCGUCC AGAAAACUCCACUGCAGAUGAAUGGAAUUGAAGAGGAUUCGGACGA ACCCCUGGAGCGCAGGCUUAGCCUCGUGCCGGAUUCAGAGCAAGGGG AGGCCAUUCUUCCCCGGAUUUCGGUGAUUUCAACCGGACCUACACUU CAGGCGAGGCGAAGGCAAUCCGUGCUCAACCUCAUGACGCAUUCGGU AAACCAGGGGCAAAACAUUCACCGCAAAACGACGGCCUCAACGAGAA AAGUGUCACUUGCACCCCAGGCGAAUUUGACUGAACUCGACAUCUAC AGCCGUAGGCUUUCGCAAGAAACCGGACUUGAGAUCAGCGAAGAAA UCAAUGAAGAAGAUUUGAAAGAGUGUUUCUUUGAUGACAUGGAAUC AAUCCCAGCGGUGACAACGUGGAACACAUACUUGCGUUACAUCACGG UGCACAAGUCCUUGAUUUUCGUCCUCAUCUGGUGUCUCGUGAUCUUU CUCGCUGAGGUCGCAGCGUCACUUGUGGUCCUCUGGCUGCUUGGUAA UACGCCCUUGCAAGACAAAGGCAAUUCUACACACUCAAGAAACAAUU CCUAUGCCGUGAUUAUCACUUCUACAAGCUCGUAUUACGUGUUUUAC AUCUACGUAGGAGUGGCCGACACUCUGCUCGCGAUGGGUUUCUUCCG AGGACUCCCACUCGUUCACACGCUUAUCACUGUCUCCAAGAUUCUCC ACCAUAAGAUGCUUCAUAGCGUACUGCAGGCUCCCAUGUCCACCUUG AAUACGCUCAAGGCGGGAGGUAUUUUGAAUCGCUUCUCAAAAGAUA UUGCAAUUUUGGAUGACCUUCUGCCCCUGACGAUCUUCGACUUCAUC CAGUUGUUGCUGAUCGUGAUUGGGGCUAUUGCAGUAGUCGCUGUCC UCCAGCCUUACAUUUUUGUCGCGACCGUUCCGGUGAUCGUGGCGUUU AUCAUGCUGCGGGCCUAUUUCUUGCAGACGUCACAGCAGCUUAAGCA ACUGGAGUCUGAAGGGAGGUCGCCUAUCUUUACGCAUCUUGUGACCA GUUUGAAGGGAUUGUGGACGUUGCGCGCCUUUGGCAGGCAGCCCUAC UUUGAAACACUGUUCCACAAAGCGCUGAAUCUCCAUACGGCAAAUUG GUUUUUGUAUUUGAGUACCCUCCGAUGGUUUCAGAUGCGCAUUGAG AUGAUUUUUGUGAUCUUCUUUAUCGCGGUGACUUUUAUCUCCAUCU UGACCACGGGAGAGGGCGAGGGACGGGUCGGUAUUAUCCUGACACUC Human CFTR Protein Sequence GCCAUGAACAUUAUGAGCACUUUGCAGUGGGCAGUGAACAGCUCGA UUGAUGUGGAUAGCCUGAUGAGGUCCGUUUCGAGGGUCUUUAAGUU CAUCGACAUGCCGACGGAGGGAAAGCCCACAAAAAGUACGAAACCCU AUAAGAAUGGGCAAUUGAGUAAGGUAAUGAUCAUCGAGAACAGUCA CGUGAAGAAGGAUGACAUCUGGCCUAGCGGGGGUCAGAUGACCGUG AAGGACCUGACGGCAAAAUACACCGAGGGAGGGAACGCAAUCCUUGA AAACAUCUCGUUCAGCAUUAGCCCCGGUCAGCGUGUGGGGUUGCUCG GGAGGACCGGGUCAGGAAAAUCGACGUUGCUGUCGGCCUUCUUGAG ACUUCUGAAUACAGAGGGUGAGAUCCAGAUCGACGGCGUUUCGUGG GAUAGCAUCACCUUGCAGCAGUGGCGGAAAGCGUUUGGAGUAAUCCC CCAAAAGGUCUUUAUCUUUAGCGGAACCUUCCGAAAGAAUCUCGAUC CUUAUGAACAGUGGUCAGAUCAAGAGAUUUGGAAAGUCGCGGACGA GGUUGGCCUUCGGAGUGUAAUCGAGCAGUUUCCGGGAAAACUCGAC UUUGUCCUUGUAGAUGGGGGAUGCGUCCUGUCGCAUGGGCACAAGC AGCUCAUGUGCCUGGCGCGAUCCGUCCUCUCUAAAGCGAAAAUUCUU CUCUUGGAUGAACCUUCGGCCCAUCUGGACCCGGUAACGUAUCAGAU CAUCAGAAGGACACUUAAGCAGGCGUUUGCCGACUGCACGGUGAUUC UCUGUGAGCAUCGUAUCGAGGCCAUGCUCGAAUGCCAGCAAUUUCUU GUCAUCGAAGAGAAUAAGGUCCGCCAGUACGACUCCAUCCAGAAGCU GCUUAAUGAGAGAUCAUUGUUCCGGCAGGCGAUUUCACCAUCCGAUA GGGUGAAACUUUUUCCACACAGAAAUUCGUCGAAGUGCAAGUCCAA | ACCGCAGAUCGCGGCCUUGAAAGAAGAGACUGAAGAAGAAGUUCAA | GACACGCGUCUUUAA MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEK | LEREWDRELASKKNPKLINALRRCFFWRFMFYGIFLYLGEVTKAVQPLLL GRIIASYDPDNKEERSIATYLGIGLCLLFIVRTLLLHPAIFGLHHIGMQMRIA e |MF SLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPLQ VALLMGLIWELLQASAFCGLGFLIVLALFQAGLGRMMMKYRDQRAGKIS ERLVITSEMIENIQSVKAYCWEEAMEKMIENLRQTELKLTRKAAYVRYFN SSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQT WYDSLGAINKIQDFLQKQEYKTLEYNLTTTEVVMENVTAFWEEGFGELFE KAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIERGQLLAVAGSTG AGKTSLLMVIMGELEPSEGKIKHSGRISFCSQFSWIMPGTIKENIIFGVSYDE YRYRSVIKACQLEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKD ADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTRILVTSKMEHLKKADKI LILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNSILTETLHR FSLEGDAPVSWTETKKQSFKQTGEFGEKRKNSILNPINSIRKFSIVQKTPLQ MNGIEEDSDEPLERRLSLVPDSEQGEAILPRISVISTGPTLQARRRQSVLNL MTHSVNQGQNIHRKTTASTRKVSLAPQANLTELDIYSRRLSQETGLEISEEI NEEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAA SLVVLWLLGNTPLQDKGNSTHSRNNSYAVITSTSSYYVFYIYVGVADTLL AMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNRFSK DIAILDDLLPLTIFDFIQLLLIVIGAIAVVAVLQPYIFVATVPVIVAFIMLRAY FLQTSQQLKQLESEGRSPIFTHLVTSLKGLWTLRAFGRQPYFETLFHKALN LHTANWFLYLSTLRWFQMRIEMIFVIFFIAVTFISILTTGEGEGRVGIILTLA MNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYKNGQ LSKVMIIENSHVKKDDIWPSGGQMTVKDLTAKYTEGGNAILENISFSISPGQ RVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWDSITLQQWRKAFGVIP QKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLDFVLVD GGCVLSHGHKQLMCLARSVLSKAKILLLDEPSAHLDPVTYQIRRTLKQAF ADCTVILCEHRIEAMLECQQFLVIEENKVRQYDSIQKLLNERSLFRQAISPS . |DRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL (SEQ IDNO:3)

[0103] Additional exemplary codon optimized mRNA sequences are described below, for example, SEQ ID NO: 7 and SEQ ID NO: 8, both of which include 5' and 3' untranslated regions framing a codon-optimized hCFTR-encoding mRNA and SEQ ID NO: 27 to SEQ ID NO: 40. Engineered or Mutant CFTR Proteins

[0104] In some embodiments, a suitable mRNA sequence encodes an engineered or mutant CFTR protein. In some embodiments, the mRNA sequence may be codon optimized. In some embodiments, an engineered or mutant CFTR protein may be a modified CFTR protein containing one or more amino acid substitutions, deletions, and / or insertions that increases the activity and / or stability as compared to a wild-type or naturally-occurring CFTR protein Accordingly, in some embodiments, the modified CFTR amino acid sequence has 70%, 75%, 80%, 85%, 90%, 95, 98%, 99% or more identity to wild type CFTR. Furthermore, in some embodiments, the modified CFTR is encoded by an mRNA that has 70%, 75%, 80%, 85%, 90%, 95, 98%, 99% or more identity to an mRNA that encodes a wild type CFTR protein. Activated CFTR Protein Mutations

[0105] CFTR is a single polypeptide containing an N-terminal lasso motif, two transmembrane domains (TMDs), and two nucleotide-binding domains (NBDs). Distinct from other ATP-binding cassette (ABC) transporters, CFTR also contains an ~200-residue cytoplasmic regulatory (R) domain that regulates the activity of CFTR. The activity of CFTR is regulated by protein kinase A-dependent phosphorylation and ATP. The R domain contains 19 predicted sites for protein kinase A (PKA); up to six have been found phosphorylated in vivo. Phosphorylation of the R domain increases the open probability of the CFTR channel and also stimulates ATP hydrolysis by CFTR. Additionally, eliminating ATP hydrolysis prolongs the lifetime of the open CFTR channel (activated CFTR), and mutating the cytosolic loops promotes CFTR activity independent of ATP.

[0106] In some embodiments, an engineered CFTR protein may contain one or more modifications that provide constitutive CFTR activity. In some embodiments, the one or more modifications provide a more CFTR activity in comparison to wild type CFTR protein activity. In particular, an engineered CFTR protein may contain one or more modifications that mimic phosphorylated residue in the R domain (R Domain Phosphomimetic mutation). In some embodiments, serine (S) or threonine (T) residues are substituted with aspartic acid (D) residues to mimic phosphorylation states. In some embodiments, serine (S) or threonine (T) residues are substituted with glutamic acid (E) residues. In some embodiments, an engineered CFTR protein may contain one or more modifications at residues $422, S660, S670, S686, T690, $700, S712, S737, S753, S768, T787, T788, S790, S795 and S813. In some embodiments, an engineered CFTR protein may contain one or more modifications that include $422D, S660D, S670D, S686D, T690D, S700D, S712D, S737D, §753D, §768D, T787D, T788D, S790D, $795D and $813D. In some embodiments, an engineered CFTR protein contains modifications of S660D, S737D, $795D and S813D. In some embodiments, an engineered CFTR protein contains modifications of S660D, S686D, S700D, S737D, S795D and S813D. In some embodiments, an engineered CFTR protein contains modifications of S660D, S686D, $700D, $712D, $737D, S768D, S795D and S813D. In some embodiments, an engineered CFTR protein contains modifications of $422D, S660D, $670D, S686D, T690D, $700D, $712D, S753D, T787D, T788D, S790D, S795D and S813D. In some embodiments, an engineered CFTR protein contains modifications of $422D, S660D, S670D, S686D, T690D, $700D, S712D, S737D, S753D, S768D, T787D, T788D, S790D, S795D and S813D.

[0107] In some embodiments, increasing the number of phosphomimetic CFTR mutations results in increased sensitivity to activation. In some embodiments, increasing the number of phosphomimetic CFTR mutations results in increased sensitivity to activation by forskolin. In some embodiments, the number of phosphomimetic CFTR mutations include 1, 2, 3,4,5,6,7,8,9,10,11, 12, 13, 14, 15,16, 17, 18, 19 or 20 amino acid substitutions.

[0108] In some embodiments, an engineered CFTR protein has low or no immunogenic potential. Accordingly, in some embodiments, an engineered CFTR protein has low immunogenic potential. In some embodiments, an engineered CFTR protein has no immunogenic potential. In some embodiments, an engineered CFTR protein does not create or enhance the potential of cytotoxic T cell lymphocyte epitopes. Accordingly, in some embodiments, an engineered CFTR protein does not create the potential of cytotoxic T cell lymphocyte epitopes. In some embodiments, an engineered CFTR protein does not enhance the potential of cytotoxic T cell lymphocyte epitopes. In some embodiments, an engineered CFTR protein does not cause increased cytotoxicity in comparison to a vehicle control.

[0109] In some embodiments, engineered, codon-optimized CFTR has increased activity in comparison to non-codon optimized CFTR.

[0110] In some embodiments, an engineered CFTR protein contains modifications at residues $660 and $737, $795 and S813. In some embodiments an engineered CFTR protein contains modifications at residues S660, S686, $700, $737, and $795. In some embodiments an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, $795, and $813. In some embodiments an engineered CFTR protein contains modifications at residues $422, $660, S670, S686, $690, $700, $712, $737, $753, S768, $787, $788, $790, $795, and $813. In some embodiments an engineered CFTR protein contains modifications at residues $422, S660, S670, S686, $690, S700, S712, $753, S787, S788, $790, $795, and S813. In some embodiments an engineered CFTR protein contains modifications at residues S660, $686, $700, $712, 8737, $768, S795, and $813. In some embodiments an engineered CFTR protein contains modifications at residue K978. In some embodiments an engineered CFTR protein contains modifications at residue E1371. In some embodiments an engineered CFTR protein contains modifications at residues $422, $660, S670, S686, S690, $700, S712, S753, S787, S788, S790, S795, S813, and E1371.In some embodiments, an engineered CFTR protein contains S660D and S737D, S795D and S813D mutations. In some embodiments an engineered CFTR protein contains S660D, S686D, S700D, S737D, and S795D mutations. In some embodiments an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, and S813D mutations. In some embodiments an engineered CFTR protein contains $422D, S660D, S670D, S686D, S690D, S700D, S712D, S737D, S753D, S768D, S787D, S788D, S790D, S795D, and S813D mutations. In some embodiments an engineered CFTR protein contains S422A, S660A, S670A, S686A, S690A, STO0A, S712A, S7T37A, S753A, S768A, ST87A, ST88A, S7T90A, S795A, and S813A mutations. In some embodiments an engineered CFTR protein contains $422D, S660D, S670D, S686D, S690D, S700D, S712D, S753D, S787D, S788D, S790D, $795D, and S813D mutations. In some embodiments an engineered CFTR protein contains S660D, S686D, S700D, S712D, S737D, S768D, S795D, and S813D mutations. In some embodiments an engineered CFTR protein contains K978C mutation. In some embodiments an engineered CFTR protein contains E1371Q mutation. In some embodiments an engineered CFTR protein contains S422A, S660A, S670A, S686A, S690A, STO0A, S712A, S753A, ST87A, S788A, S790A, S795A, S813A, and E1371Q mutations. In some embodiments an engineered CFTR protein contains $422D, $660D, S670D, S686D, S690D, S700D, $712D, $753D, S787D, S788D, S790D, §795D, S813D, and E1371Q mutations. [o111] Tn some embodiments, an engineered CFTR protein may contain one more modifications to abolish its ATPase activity (ATP Hydrolysis-Deficient mutation). Tn some embodiments, a catalytic residue in NBD2 domain of CFTR is replaced to abolish its ATPase activity. In particular embodiments, a catalytic residue is E1371. In some embodiment, an engineered CFTR protein contains an E1371Q modification.

[0112] In some embodiments, an engineered CFTR protein may contain a modification in the cytosolic loops that promote CFTR channel activity in the absence of ATP (ATP- Independent Activity mutation). In some embodiments, K978 residue in the cytosolic loop is modified to achieve constitutive activity. In some embodiments, K978 is substituted with cysteine, serine, or proline. In particular embodiments, an engineered CFTR protein contains a K978C mutation.

[0113] In some embodiments, an engineered CFTR protein may contain any combinations of modifications to achieve a CFTR protein that has greater activity as compared to wild type CFTR protein. Stable Trafficking CFTR Protein Mutations

[0114] Posttranslational modification can occur at multiple intracellular sites and modify the fate of native and damaged proteins. Both wild-type and mutant CFTR proteins undergo ubiquitination at multiple lysines in the proteins and in one or more subcellular locations. There are several potential lysines to which ubiquitin can be added in CFTR to stabilize the protein. There are additional sites on the CFTR protein that are involved in proteolysis initiation. For example, the ubiquitinated K710, K716, and K1041 residues stabilize wild-type CFTR, protecting it from proteolysis. Modifications K14R and K68R lead to increased mature band C CFTR, which can be augmented by proteasomal (but not lysosomal) inhibition, allowing trafficking to the surface. The amount of CFTR in the K1041R mutant was drastically reduced and consisted of bands A / B. The K1218R mutant increases total and cell surface CFTR, which is further accumulated by proteasomal and lysosomal inhibition.

[0115] In some embodiments, an engineered CFTR protein may contain one or more modifications that provide an increased stability for CFTR protein compared to the naturally- occurring CFTR protein (Stability mutant). Tn particular, an engineered CFTR protein may contain one or more modifications that remove an ubiquitination site in a CFTR protein. In some embodiments, an ubiquitination site is K14, K68, K710, K716, K1041, K1080 or K1218. In some embodiments, a lysine residue is substituted with an arginine residue to remove ubiquitination sites. In some embodiments an engineered CFTR protein contains one or more modifications of K14R, K68R, K710R, K716R, K1041R, K1080R, and K1218R.

[0116] In some embodiments, an engineered CFTR protein contains modifications at residue K14. In some embodiments, an engineered CFTR protein contains modifications at residue K68. In some embodiments, an engineered CFTR protein contains modifications at residues K14 and K68. In some embodiments, an engineered CFTR protein contains modifications at residue K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K68 and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K14 and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K14, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residue K710. In some embodiments, an engineered CFTR protein contains modifications at residue K716. In some embodiments, an engineered CFTR protein contains modifications at residues K710 and K716. In some embodiments, an engineered CFTR protein contains modifications at residue K1041. In some embodiments, an engineered CFTR protein contains modifications at residues K710, R716, and Kl1041.

[0117] In some embodiments, an engineered CFTR protein contains K14R mutation. In some embodiments, an engineered CFTR protein contains K68R mutation. In some embodiments, an engineered CFTR protein contains K14R and K68R mutations. In some embodiments, an engineered CFTR protein contains K1218R mutation. In some embodiments, an engineered CFTR protein contains K68R and K1218R mutations. In some embodiments, an engineered CFTR protein contains K14R and K1218R mutations. In some embodiments, an engineered CFTR protein contains K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains K710R mutation. In some embodiments, an engineered CFTR protein contains K716R mutation. In some embodiments, an engineered CFTR protein contains K710R and K716R mutations. In some embodiments, an engineered CFTR protein contains K1041R mutation. In some embodiments, an engineered CFTR protein contains K710R, R716R, and K1041R mutations.

[0118] In some embodiments, an engineered CFTR protein may contain any combination of Phosphorylated R Domain Mimic, ATP-Independent Activity, ATP Hydrolysis-Deficient, and Stability mutant modifications. In some embodiments, an engineered CFTR protein comprises one or more mutations at residues K14, K68, S422, S660, S670, $686, $700, K710, $712, K716, S737, 8753, S768, T787, T786, S790, $795, $813, K978, K1041, K1080, K1218, E1371, and combinations thereof. In particular embodiments, an engineered CFTR protein comprises a combination of mutations listed in Table 2.

[0119] In some embodiments, an engineered CFTR protein contains modifications at residues S660, $737, $795, $813, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S737, $795, $813, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues $660, $737, $795, $813, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S737, S795, S813, K14, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S737, S795, S813, K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S737, S795, S813, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S737, S795, S813, K14, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, 8795, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, S795, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, $795, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, S795, K14, and K68. In some embodiments, an engineered CFTR. protein contains modifications at residues S660, S686, S700, $737, $795, K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, S795, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, $795, K14, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K978, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues K978, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues K978 and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K978, K14, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues K978, K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K978, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues K978, K14, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, K14, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, K14, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues E1371,K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues E1371, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, $700, $737, $795, and E1371. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, 8795, E1371, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, $795, E1371, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, 8795, E1371, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, S795, E1371, K14, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, 8795, E1371, K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, $700, S737, $795, E1371, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, $700, S737, S795, E1371, K14, K68, and K1218. Tn some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, $737, $795, K978, E1371, and K14. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, S700, S737, $795, K978, E1371, and K68. Tn some embodiments, an engineered CFTR protein contains modifications at residues S660, $686, S700, $737, S795, K978, E1371, and K1218. Tn some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, $700, S737, $795, K978, E1371, K14, and K68. In some embodiments, an engineered CFTR protein contains modifications at residues S660, $686, $700, $737, $795, K978, E1371, K14, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues $660, S686, S700, $737, S795, K978, E1371, K68, and K1218. In some embodiments, an engineered CFTR protein contains modifications at residues S660, S686, $700, $737, $795, E1371, K14, K68, K978, and K1218.

[0120] In some embodiments, an engineered CFTR protein contains S660D, S73D7, $795D, $813D, and K14R mutations. In some embodiments, an engineered CFTR protein contains S660D, S737D, $795D, S813D, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S737D, S795D, S813D, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, $737D, S795D, S813D, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S737D, $795D, S813D, K14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S737D, $795D, S813D, K68R, and K1218R mutations. In some embodiments an engineered CFTR protein contains S660D, S737D, S795D, S813D. K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, $737D, $795D, and K14R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, $737D, $795D, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, §737D, S795D, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, KI14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains K978C, and K14R mutations. In some embodiments, an engineered CFTR protein contains K978C, and K68R mutations. In some embodiments, an engineered CFTR protein contains K978C and K1218R mutations. In some embodiments, an engineered CFTR protein contains K978C, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains K978C, KI14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains K978C, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains K978C, K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, and K14R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, and K68R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, and K1218R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, K14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains E1371Q, K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, E1371Q, and K14R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, $737D, $795D, E1371Q, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, $795D, E1371Q, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, E1371Q, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, E1371Q, K14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, E1371D, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, $795D, and E1371Q mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, $737D, S795D, K978C, E1371Q, and K14R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, $737D, S795D, K978C, E1371Q, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, $700D, $737D, $795D, K978C, E1371Q, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, §795D, K978C, E1371Q, K14R, and K68R mutations. In some embodiments, an engineered CFTR protein contains S660D, $686D, S700D, $737D, §795D, K978C, E1371Q, K14R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, K978C, E1371Q, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains S660D, S686D, S700D, S737D, S795D, E1371Q, K14R, K68R, and K1218R mutations. In some embodiments, an engineered CFTR protein contains $660D, S686D, S700D, S737D, $795D. E1371Q, K14R, K68R, K978C, and K1218R mutations. Table 2. Engineered CFTR Proteins Activated CFTR Mutation Stable CFTR Mutation | Engineered | S660 | S686 | S700 | $737 | S795 | S813 | K978 | E1371 | K14 | Ko68 | K1218 | CFTR A B D D D D BE le wt Bp tee gg Eee cc DD DD D DD D C E Q F R G R H - - . - . - - RR R J R R K - - . - . - - R R L R R R M D D D D R R R N DD D DD DD DD - - ~~ R RR 0 C R R R P Q R R R Q D D D D D D Q R D D D D D D Q R R R S D D D D D D C Q R R R T X X X X X X X X X X X *X = any amino acid

[0121] In some embodiments, an engineered CFTR protein comprises S660D, S737D, S795D and S813D mutations. In some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99% or more homologous to SEQ ID NO: 41. In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical SEQ ID NO: 45.

[0122] In some embodiments, an engineered CFTR protein comprises S660D, S686D, S700D, S737D, S795D and S813D mutations. In some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 42, In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical SEQ ID NO: 46.

[0123] In some embodiments, an engineered CFTR protein comprises K978C mutation. In some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%. 97%, 98%, 99% or more homologous to SEQ ID NO: 43. In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical SEQ ID NO: 47.

[0124] In some embodiments, an engineered CFTR protein comprises E1371Q mutation. In some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%. 97%, 98%, 99% or more homologous to SEQ ID NO: 44. In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 835%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical SEQ ID NO: 48. Table 3. Exemplary Engineered CFTR Protein Sequences SEQ ID NO:41 MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEK ) | LEREWDRELASKKNPKLINALRRCFFWRFMFYGIFLYLGEVTKAVQPLLL GRIIASYDPDNKEERSIATYLGIGLCLLFIVRTLLLHPAIFGLHHIGMQMRIA MFSLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPLQ VALLMGLIWELLQASAFCGLGFLIVLALFQAGLGRMMMKYRDQRAGKIS ERLVITSEMIENIQSVKAYCWEEAMEKMIENLRQTELKL TRKAAYVRYEN SSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQT WYDSLGAINKIQDFLQKQEYKTLEYNLTTTEVVMENVTAFWEEGFGELFE KAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIERGQLLAVAGSTG AGKTSLLMVIMGELEPSEGKIKHSGRISFCSQFSWIMPGTIKENIIFGVSYDE YRYRSVIKACQLEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKD ADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTRILVTSKMEHLKKADKI LILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNDILTETLHR FSLEGDAPVSWTETKKQSFKQTGEFGEKRKNSILNPINSIRKFSIVQKTPLQ MNGIEEDSDEPLERRLDLVPDSEQGEAILPRISVISTGPTLQARRRQSVLNL MTHSVNQGQNIHRKTTASTRKVDLAPQANLTELDIYSRRLDQETGLEISEET NEEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAA SLVVLWLLGNTPLQDKGNSTHSRNNSYAVIITSTSSYYVFYIYVGVADTLL AMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNRFSK DIAILDDLLPLTIFDFIQLLLIVIGAIAVVAVLQPYIFVATVPVIVAFIMLRAY FLQTSQQLKQLESEGRSPIFTHLVTSLKGLWTLRAFGRQPYFETLFHK ALNLHTANWFLYLSTLRWFQMRIEMIFVIFFIAV TFISILTTGEGEGRVGIIL TLAMNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYK NGQLSKVMIIENSHVKKDDIWPSGGQMTVKDLTAKY TEGGNAILENISFSI SPGQRVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWDSITLQQWRKA FGVIPQKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLD FVLVDGGCVLSHGHKQLMCLARSVLSKAKILLLDEPSAHLDPVTYQIIRRT LKQAFADCTVILCEHRIEAMLECQQFLVIEENKVRQYDSIQKLLNERSLFR QAISPSDRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL (SEQ 1D NO: 41) SEQ ID NO:42 MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEK ) | LEREWDRELASKKNPKLINALRRCFFWRFMFYGIFLYLGEVTKAVQPLLL GRIIASYDPDNKEERSIATYLGIGLCLLFIVRTLLLHPAIFGLHHIGMQMRIA MFSLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPLQ VALLMGLIWELLQASAFCGLGFLIVLALFQAGLGRMMMKYRDQRAGKIS ERLVITSEMIENIQSVKAY CWEEAMEKMIENLRQTELKLTRKAAYVRYFN SSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQT WYDSLGAINKIQDFLQKQEYKTLEYNLTTTEVVMENVTAFWEEGFGELFE KAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIERGQLLAVAGSTG AGKTSLLMVIMGELEPSEGKIKHSGRISFCSQFSWIMPGTIKENIIFGVSYDE YRYRSVIKACQLEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKD ADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTRILVTSKMEHLKKADKI LILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNDILTETLHR FSLEGDAPVSWTETKKQDFKQTGEFGEKRKNDILNPINSIRKFSIVQKTPLQ MNGIEEDSDEPLERRLDLVPDSEQGEAILPRISVISTGPTLQARRRQSVLNL MTHSVNQGQNIHRKTTASTRKVDLAPQANLTELDIY SRRLDQETGLEISEEI NEEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAA SLVVLWLLGNTPLQDKGNSTHSRNNSYAVITSTSSYYVFYIYVGVADTLL AMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNRFSK DIAILDDLLPLTIFDFIQLLLIVIGAIAVVAVLQPYIFVATVPVIVAFIMLRAY FLQTSQQLKQLESEGRSPIFTHLVTSLK GLWTLRAFGRQPYFETLFHKALN LHTANWFLYLSTLRWFQMRIEMIFVIFFIAVTFISILTTGEGEGRVGIILTLA MNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYKNGQ LSKVMIIENSHVKKDDIWPSGGQMTVKDLTAKY TEGGNAILENISFSISPGQ RVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWDSITLQQWRKAFGVIP QKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLDFVLVD GGCVLSHGHKQLMCLARSVLSKAKILLLDEPSAHLDPVTYQIRRTLKQAF ADCTVILCEHRIEAMLECQQFLVIEENKVRQYDSIQKLLNERSLFRQAISPS DRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL (SEQ ID NO: 42) NO:43 MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEK SEQID |LEREWDRELASKKNPKLINALRRCFFWRFMFYGIFLYLGEVTKAVQPLLL NO:43 | GRIIASYDPDNKEERSIAIYLGIGLCLLFIVRTLLLHPAIFGLHHIGMQMRIA MFSLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPL! VALLMGLIWELLQASAFCGLGFLIVLALFQAGLGRMMMKYRDQRAGKIS ERLVITSEMIENIQSVKAY CWEEAMEKMIENLRQTELKLTRKAAYVRYFN SSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQT WYDSLGAINKIQDFLQKQEYKTLEYNLTTTEVVMENV TAFWEEGFGELFE KAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIERGQLLAVAGSTG AGKTSLLMVIMGELEPSEGKIKHS GRISFCSQFSWIMPGTIKENIIFGVSYDE YRYRSVIKACQLEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKD ADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTRILVTSKMEHLKK ADKI LILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNSILTETLHR FSLEGDAPVSWTETKK QSFKQTGEFGEKRKNSILNPINSIRKFSIVQKTPLQ MNGIEEDSDEPLERRLSLVPDSEQGEAILPRISVISTGPTLQARRRQSVLNL MTHSVNQGQNIHRK TTASTRKVSLAPQANLTELDIYSRRLSQETGLEISEEI NEEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAA SLVVLWLLGNTPLQDKGNSTHSRNNSYAVIITSTSSY YVFYIYVGVADTLL AMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNRESC DIAILDDLLPLTIFDFIQLLLIVIGAIAVVAVLQPYIFVATVPVIVAFIMLRAY FLQTSQQLKQLESEGRSPIFTHLVTSLK GLWTLRAFGRQPYFETLFHKALN LHTANWELYLSTLRWFQMRIEMIFVIFFIAVTFISILTTGEGEGRVGIILTLA MNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYKNGQ LSKVMIIENSHVKKDDIWPSGGQMTVKDLTAKY TEGGNAILENISFSISPGQ RVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWDSITLQQWRKAFGVIP QKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLDFVLVD GGCVLSHGHKQLMCLARSVLSKAKILLLDEPS AHLDPVTYQIIRRTLKQAF ADCTVILCEHRIEAMLECQQFLVIEENKVRQYDSIQKLLNERSLFRQAISPS DRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL (SEQ ID NO: 43) MQRSPLEKASVVSKLFFSWTRPILRKGYRQRLELSDIYQIPSVDSADNLSEK SEQID | EREWDRELASKKNPKLINALRRCFFWRFMFYGIFLYLGEVTKAVQPLLL NO:44 | GRUASYDPDNKEERSIAIYLGIGLCLLFIVRTLLLHPAIFGLHHIGMQMRIA MFSLIYKKTLKLSSRVLDKISIGQLVSLLSNNLNKFDEGLALAHFVWIAPLQ VALLMGLIWELLQASAFCGLGFLIVLALFQAGLGRMMMKYRDQRAGKIS ERLVITSEMIENIQSVKAY CWEEAMEKMIENLRQTELKLTRKAAYVRYFN SSAFFFSGFFVVFLSVLPYALIKGIILRKIFTTISFCIVLRMAVTRQFPWAVQT WYDSLGAINKIQDFLQKQEYKTLEYNLTTTEVVMENVTAFWEEGFGELFE KAKQNNNNRKTSNGDDSLFFSNFSLLGTPVLKDINFKIERGQLLAVAGSTG AGKTSLLMVIMGELEPSEGKIKHSGRISFCSQF SWIMPGTIKENIIFGVSYDE YRYRSVIKACQLEEDISKFAEKDNIVLGEGGITLSGGQRARISLARAVYKD ADLYLLDSPFGYLDVLTEKEIFESCVCKLMANKTRILVTSKMEHLKKADKI LILHEGSSYFYGTFSELQNLQPDFSSKLMGCDSFDQFSAERRNSILTETLHR FSLEGDAPVSWTETKKQSFKQTGEFGEKRKNSILNPINSIRKFSIVQKTPLQ MNGIEEDSDEPLERRLSLVPDSEQGEAILPRISVISTGPTLQARRRQSVLNL MTHSVNQGQNIHRKTTASTRKVSLAPQANLTELDIYSRRLSQETGLEISEEI NEEDLKECFFDDMESIPAVTTWNTYLRYITVHKSLIFVLIWCLVIFLAEVAA SLVVLWLLGNTPLQDKGNSTHSRNNSYAVITSTSSYYVFYIYVGVADTLL AMGFFRGLPLVHTLITVSKILHHKMLHSVLQAPMSTLNTLKAGGILNRFSK DIAILDDLLPLTIFDFIQLLLIVIGATAVVAVLQPYIFVATVPVIVAFIMLRAY FLQTSQQLKQLESEGRSPIFTHLVTSLKGLWTLRAFGRQPYFETLFHKALN LHTANWFLYLSTLRWFQMRIEMIFVIFFIAVTFISILTTGEGEGRVGIILTLA MNIMSTLQWAVNSSIDVDSLMRSVSRVFKFIDMPTEGKPTKSTKPYKNGQ LSKVMIIENSHVKKDDIWPSGGQMTVKDLTAKY TEGGNAILENISFSISPGQ RVGLLGRTGSGKSTLLSAFLRLLNTEGEIQIDGVSWDSITLQQWRKAFGVIP QKVFIFSGTFRKNLDPYEQWSDQEIWKVADEVGLRSVIEQFPGKLDFVLVD GGCVLSHGHKQLMCLARSVLSKAKILLLDQPSAHLDPVTYQIIRRTLKQAF ADCTVILCEHRIEAMLECQQFLVIEENKVRQYDSIQKLLNERSLFRQAISPS DRVKLFPHRNSSKCKSKPQIAALKEETEEEVQDTRL (SEQ ID NO: 44) Table 4. Exemplary Codon Optimized mRNA Sequences Encoding an Engineered CFTR Protein SEQ ID NO:45 ATGCAACGCTCTCCTCTTGAAAAGGCCTCGGTGGTGTCCAAGCTCTTCT ) | TCTCGTGGACTAGACCCATCCTGAGAAAGGGGTACAGACAGCGCTTGG AGCTGTCCGATATCTATCAAATCCCTTCCGTGGACTCCGCGGACAACCT GTCCGAGAAGCTCGAGAGAGAATGGGACAGAGAACTCGCCTCAAAGA AGAACCCGAAGCTGATTAATGCGCTTAGGCGGTGCTTTTTCTGGCGGTT CATGTTCTACGGCATCTTCCTCTACCTGGGAGAGGTCACCAAGGCCGTG CAGCCCCTGTTGCTGGGACGGATTATTGCCTCCTACGACCCCGACAACA AGGAAGAAAGAAGCATCGCTATCTACTTGGGCATCGGTCTGTGCCTGC TTTTCATCGTCCGGACCCTCTTGTTGCATCCTGCTATTTICGGCCTGCAT CACATTGGCATGCAGATGAGAATTGCCATGTTTTCCCTGATCTACAAGA AAACTCTGAAGCTCTCGAGCCGCGTGCTTGACAAGATTTCCATCGGCCA GCTCGTGTCCCTGCTCTCCAACAATCTGAACAAGTTCGACGAGGGCCTC GCCCTGGCCCACTTCGTGTGGATCGCCCCTCTGCAAGTGGCGCTTCTGA TGGGCCTGATCTGGGAGCTGCTGCAAGCCTCGGCATTCTGTGGGCTTGG ATTCCTGATCGTGCTGGCACTGTTCCAGGCCGGACTGGGGCGGATGAT GATGAAGTACAGGGACCAGAGAGCCGGAAAGATTTCCGAACGGCTGG TGATCACTTCGGAAATGATCGAAAACATCCAGTCAGTGAAGGCCTACT GCTGGGAAGAGGCCATGGAAAAGATGATTGAAAACCTCCGGCAAACC GAGCTGAAGCTGACCCGCAAGGCCGCTTACGTGCGCTATTTCAACTCGT CCGCTTTCTTCTTCTCCGGGTTCTTCGTGGTGTTTCTCTCCGTGCTCCCCT ACGCCCTGATTAAGGGAATCATCCTCAGGAAGATCTTCACCACCATTTC CTTCTGTATCGTGCTCCGCATGGCCGTGACCCGGCAGTTCCCATGGGCC GTGCAGACTTGGTACGACTCCCTGGGAGCCATTAACAAGATCCAGGAC TTCCTTCAAAAGCAGGAGTACAAGACCCTCGAGTACAACCTGACTACT ACCGAGGTCGTGATGGAAAACGTCACCGCCTTTTGGGAGGAGGGATTT GGCGAACTGTTCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGAC CTCGAACGGTGACGACTCCCTCTTCTTTTCAAACTTCAGCCTGCTCGGG ACGCCCGTGCTGAAGGACATTAACTTCAAGATCGAAAGAGGACAGCTC CTGGCGGTGGCCGGATCGACCGGAGCCGGAAAGACTTCCCTGCTGATG GTGATCATGGGAGAGCTTGAACCTAGCGAGGGAAAGATCAAGCACTCC GGCCGCATCAGCTTCTGTAGCCAGTTTTCCTGGATCATGCCCGGAACCA TTAAGGAAAACATCATCTTCGGCGTGTCCTACGATGAATACCGCTACCG GTCCGTGATCAAAGCCTGCCAGCTGGAAGAGGATATTTCAAAGTTCGC GGAGAAAGATAACATCGTGCTGGGCGAAGGGGGTATTACCTTGTCGGG GGGCCAGCGGGCTAGAATCTCGCTGGCCAGAGCCGTGTATAAGGACGC CGACCTGTATCTCCTGGACTCCCCCTTCGGATACCTGGACGTCCTGACC GAAAAGGAGATCTTCGAATCGTGCGTGTGCAAGCTGATGGCTAACAAG ACTCGCATCCTCGTGACCTCCAAAATGGAGCACCTGAAGAAGGCAGAC AAGATTCTGATTCTGCATGAGGGGTCCTCCTACTTTTACGGCACCTTCT CGGAGTTGCAGAACTTGCAGCCCGACTTCTCATCGAAGCTGATGGGTT GCGACAGCTTCGACCAGTTCTCCGCCGAAAGAAGGAACGATATCCTGA CGGAAACCTTGCACCGCTTCTCTTTGGAAGGCGACGCCCCTGTGTCATG GACCGAGACTAAGAAGCAGAGCTTCAAGCAGACCGGGGAATTCGGCG AAAAGAGGAAGAACAGCATCTTGAACCCCATTAACTCCATCCGCAAGT TCTCAATCGTGCAAAAGACGCCACTGCAGATGAACGGCATTGAGGAGG ACTCCGACGAACCCCTTGAGAGGCGCCTGGATCTGGTGCCGGACAGCG AGCAGGGAGAAGCCATCCTGCCTCGGATTTCCGTGATCTCCACTGGTCC GACGCTCCAAGCCCGGCGGCGGCAGTCCGTGCTGAACCTGATGACCCA CAGCGTGAACCAGGGCCAAAACATTCACCGCAAGACTACCGCATCCAC CCGGAAAGTGGATCTGGCACCTCAAGCGAATCTTACCGAGCTCGACAT CTACTCCCGGAGACTGGATCAGGAAACCGGGCTCGAAATTTCCGAAGA AATCAACGAGGAGGATCTGAAAGAGTGCTTCTTCGACGATATGGAGTC GATACCCGCCGTGACGACTTGGAACACTTATCTGCGGTACATCACTGTG CACAAGTCATTGATCTTCGTGCTGATTTGGTGCCTGGTGATTTTCCTGG CCGAGGTCGCGGCCTCACTGGTGGTGCTCTGGCTGTTGGGAAACACGC CTCTGCAAGACAAGGGAAACTCCACGCACTCGAGAAACAACAGCTATG CCGTGATTATCACTTCCACCTCCTCTTATTACGTGTTCTACATCTACGTC GGAGTGGCGGATACCCTGCTCGCGATGGGTTTCTTCAGAGGACTGCCG CTGGTCCACACCTTGATCACCGTCAGCAAGATTCTTCACCACAAGATGT TGCATAGCGTGCTGCAGGCCCCCATGTCCACCCTCAACACTCTGAAGGC CGGAGGCATTCTGAACAGATTCTCCAAGGACATCGCTATCCTGGACGA TCTCCTGCCGCTTACCATCTTTGACTTCATCCAGCTGCTGCTGATCGTGA TTGGAGCAATCGCAGTGGTGGCGGTGCTGCAGCCTTACATTTTCGTGGC CACTGTGCCGGTCATTGTGGCGTTCATCATGCTGCGGGCCTACTTCCTC CAAACCAGCCAGCAGCTGAAGCAACTGGAATCCGAGGGACGATCCCCC ATCTTCACTCACCTTGTGACGTCGTTGAAGGGACTGTGGACCCTCCGGG CTTTCGGACGGCAGCCCTACTTCGAAACCCTCTTCCACAAGGCCCTGAA CCTCCACACCGCCAATTGGTTCCTGTACCTGTCCACCCTGCGGTGGTTC CAGATGCGCATCGAGATGATTTTCGTCATCTTCTTCATCGCGGTCACAT TCATCAGCATCCTGACTACCGGAGAGGGAGAGGGACGGGTCGGAATAA TCCTGACCCTCGCCATGAACATTATGAGCACCCTGCAGTGGGCAGTGA ACAGCTCGATCGACGTGGACAGCCTGATGCGAAGCGTCAGCCGCGTGT TCAAGTTCATCGACATGCCTACTGAGGGAAAACCCACTAAGTCCACTA AGCCCTACAAAAATGGCCAGCTGAGCAAGGTCATGATCATCGAAAACT CCCACGTGAAGAAGGACGATATTTGGCCCTCCGGAGGTCAAATGACCG TGAAGGACCTGACCGCAAAGTACACCGAGGGAGGAAACGCCATTCTCG AAAACATCAGCTTCTCCATTTCGCCGGGACAGCGGGTCGGCCTTCTCGG GCGGACCGGTTCCGGGAAGTCAACTCTGCTGTCGGCTTTCCTCCGGCTG CTGAATACCGAGGGGGAAATCCAAATTGACGGCGTGTCTTGGGATTCC ATTACTCTGCAGCAGTGGCGGAAGGCCTTCGGCGTGATCCCCCAGAAG GTGTTCATCTTCTCGGGTACCTTCCGGAAGAACCTGGATCCTTACGAGC AGTGGAGCGACCAAGAAATCTGGAAGGTCGCCGACGAGGTCGGCCTGC GCTCCGTGATTGAACAATTTCCTGGAAAGCTGGACTTCGTGCTCGTCGA CGGGGGATGTGTCCTGTCGCACGGACATAAGCAGCTCATGTGCCTCGC ACGGTCCGTGCTCTCCAAGGCCAAGATTCTGCTGCTGGACGAACCTTCG GCCCACCTGGATCCGGTCACCTACCAGATCATCAGGAGGACCCTGAAG CAGGCCTTTGCCGATTGCACCGTGATTCTCTGCGAGCACCGCATCGAGG CCATGCTGGAGTGCCAGCAGTTCCTGGTCATCGAGGAGAACAAGGTCC GCCAATACGACTCCATTCAAAAGCTCCTCAACGAGCGGTCGCTGTTCA GACAAGCTATTTCACCGTCCGATAGAGTGAAGCTCTTCCCGCATCGGA ACAGCTCAAAGTGCAAATCGAAGCCGCAGATCGCAGCCTTGAAGGAAG AGACTGAGGAAGAGGTGCAGGACACCCGGCTTTAA (SEQ ID NO: 45) ATGCAACGCTCTCCTCTTGAAAAGGCCTCGGTGGTGTCCAAGCTCTTCT SEQID | TCTCGTGGACTAGACCCATCCTGAGAAAGGGGTACAGACAGCGCTTGG NO:46 | AGCTGTCCGATATCTATCAAATCCCTTCCGTGGACTCCGCGGACAACCT GTCCGAGAAGCTCGAGAGAGAATGGGACAGAGAACTCGCCTCAAAGA AGAACCCGAAGCTGATTAATGCGCTTAGGCGGTGCTTTTTCTGGCGGTT CATGTTCTACGGCATCTTCCTCTACCTGGGAGAGGTCACCAAGGCCGTG CAGCCCCTGTTGCTGGGACGGATTATTGCCTCCTACGACCCCGACAACA AGGAAGAAAGAAGCATCGCTATCTACTTGGGCATCGGTCTGTGCCTGC TTTTCATCGTCCGGACCCTCTTGTTGCATCCTGCTATTTTCGGCCTGCAT CACATTGGCATGCAGATGAGAATTGCCATGTTTTCCCTGATCTACAAGA AAACTCTGAAGCTCTCGAGCCGCGTGCTTGACAAGATTTCCATCGGCCA GCTCGTGTCCCTGCTCTCCAACAATCTGAACAAGTTCGACGAGGGCCTC GCCCTGGCCCACTTCGTGTGGATCGCCCCTCTGCAAGTGGCGCTTCTGA TGGGCCTGATCTGGGAGCTGCTGCAAGCCTCGGCATTCTGTGGGCTTGG ATTCCTGATCGTGCTGGCACTGTTCCAGGCCGGACTGGGGCGGATGAT GATGAAGTACAGGGACCAGAGAGCCGGAAAGATTTCCGAACGGCTGG TGATCACTTCGGAAATGATCGAAAACATCCAGTCAGTGAAGGCCTACT GCTGGGAAGAGGCCATGGAAAAGATGATTGAAAACCTCCGGCAAACC GAGCTGAAGCTGACCCGCAAGGCCGCTTACGTGCGCTATTTCAACTCGT CCGCTTTCTTCTTCTCCGGGTTCTTCGTGGTGTTITCTCTCCGTGCTCCCCT ACGCCCTGATTAAGGGAATCATCCTCAGGAAGATCTTCACCACCATTTC CTTCTGTATCGTGCTCCGCATGGCCGTGACCCGGCAGTTCCCATGGGCC GTGCAGACTTGGTACGACTCCCTGGGAGCCATTAACAAGATCCAGGAC TTCCTTCAAAAGCAGGAGTACAAGACCCTCGAGTACAACCTGACTACT ACCGAGGTCGTGATGGAAAACGTCACCGCCTTTTGGGAGGAGGGATTT GGCGAACTGTTCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGAC CTCGAACGGTGACGACTCCCTCTTCTTTTCAAACTTCAGCCTGCTCGGG ACGCCCGTGCTGAAGGACATTAACTTCAAGATCGAAAGAGGACAGCTC CTGGCGGTGGCCGGATCGACCGGAGCCGGAAAGACTTCCCTGCTGATG GTGATCATGGGAGAGCTTGAACCTAGCGAGGGAAAGATCAAGCACTCC GGCCGCATCAGCTTCTGTAGCCAGTTTTCCTGGATCATGCCCGGAACCA TTAAGGAAAACATCATCTTCGGCGTGTCCTACGATGAATACCGCTACCG GTCCGTGATCAAAGCCTGCCAGCTGGAAGAGGATATTTCAAAGTTCGC GGAGAAAGATAACATCGTGCTGGGCGAAGGGGGTATTACCTTGTCGGG GGGCCAGCGGGCTAGAATCTCGCTGGCCAGAGCCGTGTATAAGGACGC CGACCTGTATCTCCTGGACTCCCCCTTCGGATACCTGGACGTCCTGACC GAAAAGGAGATCTTCGAATCGTGCGTGTGCAAGCTGATGGCTAACAAG ACTCGCATCCTCGTGACCTCCAAAATGGAGCACCTGAAGAAGGCAGAC AAGATTCTGATTCTGCATGAGGGGTCCTCCTACTTTTACGGCACCTTCT CGGAGTTGCAGAACTTGCAGCCCGACTTCTCATCGAAGCTGATGGGTT GCGACAGCTTCGACCAGTTCTCCGCCGAAAGAAGGAACGATATCCTGA CGGAAACCTTGCACCGCTTCTCTTTGGAAGGCGACGCCCCTGTGTCATG GACCGAGACTAAGAAGCAGGATTTCAAGCAGACCGGGGAATTCGGCG AAAAGAGGAAGAACGACATCTTGAACCCCATTAACTCCATCCGCAAGT TCTCAATCGTGCAAAAGACGCCACTGCAGATGAACGGCATTGAGGAGG ACTCCGACGAACCCCTTGAGAGGCGCCTGGATCTGGTGCCGGACAGCG AGCAGGGAGAAGCCATCCTGCCTCGGATTTCCGTGATCTCCACTGGTCC GACGCTCCAAGCCCGGCGGCGGCAGTCCGTGCTGAACCTGATGACCCA CAGCGTGAACCAGGGCCAAAACATTCACCGCAAGACTACCGCATCCAC CCGGAAAGTGGATCTGGCACCTCAAGCGAATCTTACCGAGCTCGACAT CTACTCCCGGAGACTGGATCAGGAAACCGGGCTCGAAATTTCCGAAGA AATCAACGAGGAGGATCTGAAAGAGTGCTTCTTCGACGATATGGAGTC GATACCCGCCGTGACGACTTGGAACACTTATCTGCGGTACATCACTGTG CACAAGTCATTGATCTTCGTGCTGATTTGGTGCCTGGTGATTTTCCTGG CCGAGGTCGCGGCCTCACTGGTGGTGCTCTGGCTGTTGGGAAACACGC CTCTGCAAGACAAGGGAAACTCCACGCACTCGAGAAACAACAGCTATG CCGTGATTATCACTTCCACCTCCTCTTATTACGTGTTCTACATCTACGTC GGAGTGGCGGATACCCTGCTCGCGATGGGTTTCTTCAGAGGACTGCCG CTGGTCCACACCTTGATCACCGTCAGCAAGATTCTTCACCACAAGATGT TGCATAGCGTGCTGCAGGCCCCCATGTCCACCCTCAACACTCTGAAGGC CGGAGGCATTCTGAACAGATTCTCCAAGGACATCGCTATCCTGGACGA TCTCCTGCCGCTTACCATCTTTGACTTCATCCAGCTGCTGCTGATCGTGA TTGGAGCAATCGCAGTGGTGGCGGTGCTGCAGCCTTACATTTTCGTGGC CACTGTGCCGGTCATTGTGGCGTTCATCATGCTGCGGGCCTACTTCCTC CAAACCAGCCAGCAGCTGAAGCAACTGGAATCCGAGGGACGATCCCCC ATCTTCACTCACCTTGTGACGTCGTTGAAGGGACTGTGGACCCTCCGGG CTTTCGGACGGCAGCCCTACTTCGAAACCCTCTTCCACAAGGCCCTGAA CCTCCACACCGCCAATTGGTTCCTGTACCTGTCCACCCTGCGGTGGTTC CAGATGCGCATCGAGATGATTTTCGTCATCTTCTTCATCGCGGTCACAT TCATCAGCATCCTGACTACCGGAGAGGGAGAGGGACGGGTCGGAATAA TCCTGACCCTCGCCATGAACATTATGAGCACCCTGCAGTGGGCAGTGA ACAGCTCGATCGACGTGGACAGCCTGATGCGAAGCGTCAGCCGCGTGT TCAAGTTCATCGACATGCCTACTGAGGGAAAACCCACTAAGTCCACTA AGCCCTACAAAAATGGCCAGCTGAGCAAGGTCATGATCATCGAAAACT CCCACGTGAAGAAGGACGATATTTGGCCCTCCGGAGGTCAAATGACCG TGAAGGACCTGACCGCAAAGTACACCGAGGGAGGAAACGCCATTCTCG AAAACATCAGCTTCTCCATTTCGCCGGGACAGCGGGTCGGCCTTCTCGG GCGGACCGGTTCCGGGAAGTCAACTCTGCTGTCGGCTTTCCTCCGGCTG CTGAATACCGAGGGGGAAATCCAAATTGACGGCGTGTCTTGGGATTCC ATTACTCTGCAGCAGTGGCGGAAGGCCTTCGGCGTGATCCCCCAGAAG GTGTTCATCTTCTCGGGTACCTTCCGGAAGAACCTGGATCCTTACGAGC AGTGGAGCGACCAAGAAATCTGGAAGGTCGCCGACGAGGTCGGCCTGC GCTCCGTGATTGAACAATTTCCTGGAAAGCTGGACTTCGTGCTCGTCGA CGGGGGATGTGTCCTGTCGCACGGACATAAGCAGCTCATGTGCCTCGC ACGGTCCGTGCTCTCCAAGGCCAAGATTCTGCTGCTGGACGAACCITCG GCCCACCTGGATCCGGTCACCTACCAGATCATCAGGAGGACCCTGAAG CAGGCCTTTGCCGATTGCACCGTGATTCTCTGCGAGCACCGCATCGAGG CCATGCTGGAGTGCCAGCAGTTCCTGGTCATCGAGGAGAACAAGGTCC GCCAATACGACTCCATTCAAAAGCTCCTCAACGAGCGGTCGCTGTTCA GACAAGCTATTTCACCGTCCGATAGAGTGAAGCTCTTCCCGCATCGGA ACAGCTCAAAGTGCAAATCGAAGCCGCAGATCGCAGCCTTGAAGGAAG AGACTGAGGAAGAGGTGCAGGACACCCGGCTTTAA (SEQ ID NO: 46) ATGCAACGCTCTCCTCTTGAAAAGGCCTCGGTGGTGTCCAAGCTCTTCT SEQID | TCTCGTGGACTAGACCCATCCTGAGAAAGGGGTACAGACAGCGCTTGG NO:47 | AGCTGTCCGATATCTATCAAATCCCTTCCGTGGACTCCGCGGACAACCT GTCCGAGAAGCTCGAGAGAGAATGGGACAGAGAACTCGCCTCAAAGA AGAACCCGAAGCTGATTAATGCGCTTAGGCGGTGCTTTTTCTGGCGGTT CATGTTCTACGGCATCTTCCTCTACCTGGGAGAGGTCACCAAGGCCGTG CAGCCCCTGTTGCTGGGACGGATTATTGCCTCCTACGACCCCGACAACA AGGAAGAAAGAAGCATCGCTATCTACTTGGGCATCGGTCTGTGCCTGC TTTTCATCGTCCGGACCCTCTTGTTGCATCCTGCTATTTTCGGCCTGCAT CACATTGGCATGCAGATGAGAATTGCCATGTTTTCCCTGATCTACAAGA AAACTCTGAAGCTCTCGAGCCGCGTGCTTGACAAGATTTCCATCGGCCA GCTCGTGTCCCTGCTCTCCAACAATCTGAACAAGTTCGACGAGGGCCTC GCCCTGGCCCACTTCGTGTGGATCGCCCCTCTGCAAGTGGCGCTTCTGA TGGGCCTGATCTGGGAGCTGCTGCAAGCCTCGGCATTCTGTGGGCTTGG ATTCCTGATCGTGCTGGCACTGTTCCAGGCCGGACTGGGGCGGATGAT GATGAAGTACAGGGACCAGAGAGCCGGAAAGATTTCCGAACGGCTGG TGATCACTTCGGAAATGATCGAAAACATCCAGTCAGTGAAGGCCTACT GCTGGGAAGAGGCCATGGAAAAGATGATTGAAAACCTCCGGCAAACC GAGCTGAAGCTGACCCGCAAGGCCGCTTACGTGCGCTATTTCAACTCGT CCGCTTTCTTCTTCTCCGGGTTCTTCGTGGTGTTITCTCTCCGTGCTCCCCT ACGCCCTGATTAAGGGAATCATCCTCAGGAAGATCTTCACCACCATTIC CTTCTGTATCGTGCTCCGCATGGCCGTGACCCGGCAGTTCCCATGGGCC GTGCAGACTTGGTACGACTCCCTGGGAGCCATTAACAAGATCCAGGAC TTCCTTCAAAAGCAGGAGTACAAGACCCTCGAGTACAACCTGACTACT ACCGAGGTCGTGATGGAAAACGTCACCGCCTTTTGGGAGGAGGGATTT GGCGAACTGTTCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGAC CTCGAACGGTGACGACTCCCTCTTCTTTTCAAACTTCAGCCTGCTCGGG ACGCCCGTGCTGAAGGACATTAACTTCAAGATCGAAAGAGGACAGCTC CTGGCGGTGGCCGGATCGACCGGAGCCGGAAAGACTTCCCTGCTGATG GTGATCATGGGAGAGCTTGAACCTAGCGAGGGAAAGATCAAGCACTCC GGCCGCATCAGCTTCTGTAGCCAGTTTTCCTGGATCATGCCCGGAACCA TTAAGGAAAACATCATCTTCGGCGTGTCCTACGATGAATACCGCTACCG GTCCGTGATCAAAGCCTGCCAGCTGGAAGAGGATATTTCAAAGTTCGC GGAGAAAGATAACATCGTGCTGGGCGAAGGGGGTATTACCTTGTCGGG GGGCCAGCGGGCTAGAATCTCGCTGGCCAGAGCCGTGTATAAGGACGC CGACCTGTATCTCCTGGACTCCCCCTTCGGATACCTGGACGTCCTGACC GAAAAGGAGATCTTCGAATCGTGCGTGTGCAAGCTGATGGCTAACAAG ACTCGCATCCTCGTGACCTCCAAAATGGAGCACCTGAAGAAGGCAGAC AAGATTCTGATTCTGCATGAGGGGTCCTCCTACTTTTACGGCACCTTCT CGGAGTTGCAGAACTTGCAGCCCGACTTCTCATCGAAGCTGATGGGTT GCGACAGCTTCGACCAGTTCTCCGCCGAAAGAAGGAACTCGATCCTGA CGGAAACCTTGCACCGCTTCTCTTTGGAAGGCGACGCCCCTGTGTCATG GACCGAGACTAAGAAGCAGAGCTTCAAGCAGACCGGGGAATTCGGCG AAAAGAGGAAGAACAGCATCTTGAACCCCATTAACTCCATCCGCAAGT TCTCAATCGTGCAAAAGACGCCACTGCAGATGAACGGCATTGAGGAGG ACTCCGACGAACCCCTTGAGAGGCGCCTGTCCCTGGTGCCGGACAGCG AGCAGGGAGAAGCCATCCTGCCTCGGATTTCCGTGATCTCCACTGGTCC GACGCTCCAAGCCCGGCGGCGGCAGTCCGTGCTGAACCTGATGACCCA CAGCGTGAACCAGGGCCAAAACATTCACCGCAAGACTACCGCATCCAC CCGGAAAGTGTCCCTGGCACCTCAAGCGAATCTTACCGAGCTCGACAT CTACTCCCGGAGACTGTCGCAGGAAACCGGGCTCGAAATTTCCGAAGA AATCAACGAGGAGGATCTGAAAGAGTGCTTCTTCGACGATATGGAGTC GATACCCGCCGTGACGACTTGGAACACTTATCTGCGGTACATCACTGTG CACAAGTCATTGATCTTCGTGCTGATTTGGTGCCTGGTGATTTTCCTGG CCGAGGTCGCGGCCTCACTGGTGGTGCTCTGGCTGTTGGGAAACACGC CTCTGCAAGACAAGGGAAACTCCACGCACTCGAGAAACAACAGCTATG CCGTGATTATCACTTCCACCTCCTCTTATTACGTGTTCTACATCTACGTC GGAGTGGCGGATACCCTGCTCGCGATGGGTTTCTTCAGAGGACTGCCG CTGGTCCACACCTTGATCACCGTCAGCAAGATTCTTCACCACAAGATGT TGCATAGCGTGCTGCAGGCCCCCATGTCCACCCTCAACACTCTGAAGGC CGGAGGCATTCTGAACAGATTCTCCTGCGACATCGCTATCCTGGACGAT CTCCTGCCGCTTACCATCTTTGACTTCATCCAGCTGCTGCTGATCGTGAT TGGAGCAATCGCAGTGGTGGCGGTGCTGCAGCCTTACATTTTCGTGGCC ACTGTGCCGGTCATTGTGGCGTTCATCATGCTGCGGGCCTACTTCCTCC AAACCAGCCAGCAGCTGAAGCAACTGGAATCCGAGGGACGATCCCCCA TCTTCACTCACCTTGTGACGTCGTTGAAGGGACTGTGGACCCTCCGGGC TTTCGGACGGCAGCCCTACTTCGAAACCCTCTTCCACAAGGCCCTGAAC CTCCACACCGCCAATTGGTTCCTGTACCTGTCCACCCTGCGGTGGTTCC AGATGCGCATCGAGATGATTTTCGTCATCTTCTTCATCGCGGTCACATT CATCAGCATCCTGACTACCGGAGAGGGAGAGGGACGGGTCGGAATAAT CCTGACCCTCGCCATGAACATTATGAGCACCCTGCAGTGGGCAGTGAA CAGCTCGATCGACGTGGACAGCCTGATGCGAAGCGTCAGCCGCGTGTT CAAGTTCATCGACATGCCTACTGAGGGAAAACCCACTAAGTCCACTAA GCCCTACAAAAATGGCCAGCTGAGCAAGGTCATGATCATCGAAAACTC CCACGTGAAGAAGGACGATATTTGGCCCTCCGGAGGTCAAATGACCGT GAAGGACCTGACCGCAAAGTACACCGAGGGAGGAAACGCCATTCTCG AAAACATCAGCTTCTCCATTTCGCCGGGACAGCGGGTCGGCCTTCTCGG GCGGACCGGTTCCGGGAAGTCAACTCTGCTGTCGGCTTTCCTCCGGCTG CTGAATACCGAGGGGGAAATCCAAATTGACGGCGTGTCTTGGGATTCC ATTACTCTGCAGCAGTGGCGGAAGGCCTTCGGCGTGATCCCCCAGAAG GTGTTCATCTTCTCGGGTACCTTCCGGAAGAACCTGGATCCTTACGAGC AGTGGAGCGACCAAGAAATCTGGAAGGTCGCCGACGAGGTCGGCCTGC GCTCCGTGATTGAACAATTTCCTGGAAAGCTGGACTTCGTGCTCGTCGA CGGGGGATGTGTCCTGTCGCACGGACATAAGCAGCTCATGTGCCTCGC ACGGTCCGTGCTCTCCAAGGCCAAGATTCTGCTGCTGGACGAACCITCG GCCCACCTGGATCCGGTCACCTACCAGATCATCAGGAGGACCCTGAAG CAGGCCTTTGCCGATTGCACCGTGATTCTCTGCGAGCACCGCATCGAGG CCATGCTGGAGTGCCAGCAGTTCCTGGTCATCGAGGAGAACAAGGTCC GCCAATACGACTCCATTCAAAAGCTCCTCAACGAGCGGTCGCTGTTCA GACAAGCTATTTCACCGTCCGATAGAGTGAAGCTCTTCCCGCATCGGA ACAGCTCAAAGTGCAAATCGAAGCCGCAGATCGCAGCCTTGAAGGAAG AGACTGAGGAAGAGGTGCAGGACACCCGGCTTTAA (SEQ ID NO: 47) ATGCAACGCTCTCCTCTTGAAAAGGCCTCGGTGGTGTCCAAGCTCTTCT SEQID | TCTCGTGGACTAGACCCATCCTGAGAAAGGGGTACAGACAGCGCTTGG NO:48 | AGCTGTCCGATATCTATCAAATCCCTTCCGTGGACTCCGCGGACAACCT GTCCGAGAAGCTCGAGAGAGAATGGGACAGAGAACTCGCCTCAAAGA AGAACCCGAAGCTGATTAATGCGCTTAGGCGGTGCTTTTTCTGGCGGTT CATGTTCTACGGCATCTTCCTCTACCTGGGAGAGGTCACCAAGGCCGTG CAGCCCCTGTTGCTGGGACGGATTATTGCCTCCTACGACCCCGACAACA AGGAAGAAAGAAGCATCGCTATCTACTTGGGCATCGGTCTGTGCCTGC TTTTCATCGTCCGGACCCTCTTGTTGCATCCTGCTATTTTCGGCCTGCAT CACATTGGCATGCAGATGAGAATTGCCATGTTTTCCCTGATCTACAAGA AAACTCTGAAGCTCTCGAGCCGCGTGCTTGACAAGATTTCCATCGGCCA GCTCGTGTCCCTGCTCTCCAACAATCTGAACAAGTTCGACGAGGGCCTC GCCCTGGCCCACTTCGTGTGGATCGCCCCTCTGCAAGTGGCGCTTCTGA TGGGCCTGATCTGGGAGCTGCTGCAAGCCTCGGCATTCTGTGGGCTTGG ATTCCTGATCGTGCTGGCACTGTTCCAGGCCGGACTGGGGCGGATGAT GATGAAGTACAGGGACCAGAGAGCCGGAAAGATTTCCGAACGGCTGG TGATCACTTCGGAAATGATCGAAAACATCCAGTCAGTGAAGGCCTACT GCTGGGAAGAGGCCATGGAAAAGATGATTGAAAACCTCCGGCAAACC GAGCTGAAGCTGACCCGCAAGGCCGCTTACGTGCGCTATTITCAACTCGT CCGCTTTCTTICTTCTCCGGGTTCTTCGTGGTGTTTCTCTCCGTGCTCCCCT ACGCCCTGATTAAGGGAATCATCCTCAGGAAGATCTTCACCACCATTIC CTTCTGTATCGTGCTCCGCATGGCCGTGACCCGGCAGTTCCCATGGGCC GTGCAGACTTGGTACGACTCCCTGGGAGCCATTAACAAGATCCAGGAC TTCCTTCAAAAGCAGGAGTACAAGACCCTCGAGTACAACCTGACTACT ACCGAGGTCGTGATGGAAAACGTCACCGCCTTTTGGGAGGAGGGATTT GGCGAACTGTTCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGAC CTCGAACGGTGACGACTCCCTCTTCTTTTCAAACTTCAGCCTGCTCGGG ACGCCCGTGCTGAAGGACATTAACTTCAAGATCGAAAGAGGACAGCTC CTGGCGGTGGCCGGATCGACCGGAGCCGGAAAGACTTCCCTGCTGATG GTGATCATGGGAGAGCTTGAACCTAGCGAGGGAAAGATCAAGCACTCC GGCCGCATCAGCTTCTGTAGCCAGTTTTCCTGGATCATGCCCGGAACCA TTAAGGAAAACATCATCTTCGGCGTGTCCTACGATGAATACCGCTACCG GTCCGTGATCAAAGCCTGCCAGCTGGAAGAGGATATTTCAAAGTTCGC GGAGAAAGATAACATCGTGCTGGGCGAAGGGGGTATTACCTTGTCGGG GGGCCAGCGGGCTAGAATCTCGCTGGCCAGAGCCGTGTATAAGGACGC CGACCTGTATCTCCTGGACTCCCCCTTCGGATACCTGGACGTCCTGACC GAAAAGGAGATCTTCGAATCGTGCGTGTGCAAGCTGATGGCTAACAAG ACTCGCATCCTCGTGACCTCCAAAATGGAGCACCTGAAGAAGGCAGAC AAGATTCTGATTCTGCATGAGGGGTCCTCCTACTTTTACGGCACCTTCT CGGAGTTGCAGAACTTGCAGCCCGACTTCTCATCGAAGCTGATGGGTT GCGACAGCTTCGACCAGTTCTCCGCCGAAAGAAGGAACTCGATCCTGA CGGAAACCTTGCACCGCTTCTCTTTGGAAGGCGACGCCCCTGTGTCATG GACCGAGACTAAGAAGCAGAGCTTCAAGCAGACCGGGGAATTCGGCG AAAAGAGGAAGAACAGCATCTTGAACCCCATTAACTCCATCCGCAAGT TCTCAATCGTGCAAAAGACGCCACTGCAGATGAACGGCATTGAGGAGG ACTCCGACGAACCCCTTGAGAGGCGCCTGTCCCTGGTGCCGGACAGCG AGCAGGGAGAAGCCATCCTGCCTCGGATTTCCGTGATCTCCACTGGTCC GACGCTCCAAGCCCGGCGGCGGCAGTCCGTGCTGAACCTGATGACCCA CAGCGTGAACCAGGGCCAAAACATTCACCGCAAGACTACCGCATCCAC CCGGAAAGTGTCCCTGGCACCTCAAGCGAATCTTACCGAGCTCGACAT CTACTCCCGGAGACTGTCGCAGGAAACCGGGCTCGAAATTTCCGAAGA AATCAACGAGGAGGATCTGAAAGAGTGCTTCTTCGACGATATGGAGTC GATACCCGCCGTGACGACTTGGAACACTTATCTGCGGTACATCACTGTG CACAAGTCATTGATCTTCGTGCTGATTTGGTGCCTGGTGATTTTCCTGG CCGAGGTCGCGGCCTCACTGGTGGTGCTCTGGCTGTTGGGAAACACGC CTCTGCAAGACAAGGGAAACTCCACGCACTCGAGAAACAACAGCTATG CCGTGATTATCACTTCCACCTCCTCTTATTACGTGTTCTACATCTACGTC GGAGTGGCGGATACCCTGCTCGCGATGGGTTTCTTCAGAGGACTGCCG CTGGTCCACACCTTGATCACCGTCAGCAAGATTCTTCACCACAAGATGT TGCATAGCGTGCTGCAGGCCCCCATGTCCACCCTCAACACTCTGAAGGC CGGAGGCATTCTGAACAGATTCTCCAAGGACATCGCTATCCTGGACGA TCTCCTGCCGCTTACCATCTTTGACTTCATCCAGCTGCTGCTGATCGTGA TTGGAGCAATCGCAGTGGTGGCGGTGCTGCAGCCTTACATTTTCGTGGC CACTGTGCCGGTCATTGTGGCGTTCATCATGCTGCGGGCCTACTTCCTC CAAACCAGCCAGCAGCTGAAGCAACTGGAATCCGAGGGACGATCCCCC ATCTTCACTCACCTTGTGACGTCGTTGAAGGGACTGTGGACCCTCCGGG CTTTCGGACGGCAGCCCTACTTCGAAACCCTCTTCCACAAGGCCCTGAA CCTCCACACCGCCAATTGGTTCCTGTACCTGTCCACCCTGCGGTGGTTC CAGATGCGCATCGAGATGATTTTCGTCATCTTCTTCATCGCGGTCACAT Codon hCFTR - K14R E1371Q TCATCAGCATCCTGACTACCGGAGAGGGAGAGGGACGGGTCGGAATAA TCCTGACCCTCGCCATGAACATTATGAGCACCCTGCAGTGGGCAGTGA ACAGCTCGATCGACGTGGACAGCCTGATGCGAAGCGTCAGCCGCGTGT TCAAGTTCATCGACATGCCTACTGAGGGAAAACCCACTAAGTCCACTA AGCCCTACAAAAATGGCCAGCTGAGCAAGGTCATGATCATCGAAAACT CCCACGTGAAGAAGGACGATATTTGGCCCTCCGGAGGTCAAATGACCG TGAAGGACCTGACCGCAAAGTACACCGAGGGAGGAAACGCCATTCTCG AAAACATCAGCTTCTCCATTTCGCCGGGACAGCGGGTCGGCCTTCTCGG GCGGACCGGTTCCGGGAAGTCAACTCTGCTGTCGGCTTTCCTCCGGCTG CTGAATACCGAGGGGGAAATCCAAATTGACGGCGTGTCTTGGGATTCC ATTACTCTGCAGCAGTGGCGGAAGGCCTTCGGCGTGATCCCCCAGAAG GTGTTCATCTTCTCGGGTACCTTCCGGAAGAACCTGGATCCTTACGAGC AGTGGAGCGACCAAGAAATCTGGAAGGTCGCCGACGAGGTCGGCCTGC GCTCCGTGATTGAACAATTTCCTGGAAAGCTGGACTTCGTGCTCGTCGA CGGGGGATGTGTCCTGTCGCACGGACATAAGCAGCTCATGTGCCTCGC ACGGTCCGTGCTCTCCAAGGCCAAGATTCTGCTGCTGGACCAACCTTCG GCCCACCTGGATCCGGTCACCTACCAGATCATCAGGAGGACCCTGAAG CAGGCCTTTGCCGATTGCACCGTGATTCTCTGCGAGCACCGCATCGAGG CCATGCTGGAGTGCCAGCAGTTCCTGGTCATCGAGGAGAACAAGGTCC GCCAATACGACTCCATTCAAAAGCTCCTCAACGAGCGGTCGCTGTTCA GACAAGCTATTTCACCGTCCGATAGAGTGAAGCTCTTCCCGCATCGGA ACAGCTCAAAGTGCAAATCGAAGCCGCAGATCGCAGCCTTGAAGGAAG AGACTGAGGAAGAGGTGCAGGACACCCGGCTTTAA (SEQ ID NO: 48) ATGCAGCGCTCGCCTCTGGAAAAGGCGAGCGTCGTGTCACGGCTATTIC SEQID |TTTTCTTGGACCCGGCCCATTCTCAGGAAGGGCTACAGGCAGAGGCTG NO:49 | GAGTTGAGCGACATCTATCAGATTCCTTCCGTGGACAGCGCCGACAAC CTGAGCGAGAAGCTGGAAAGGGAGTGGGACCGCGAACTGGCAAGCAA Codon | AAAGAACCCCAAGCTGATCAATGCCCTGAGAAGGTGTTTCTITTGGAG Optimized | ATTCATGTTCTACGGGATCTTTCTGTATCTGGGCGAGGTTACAAAGGCT hCFTR- | GTGCAGCCCCTGCTGCTCGGCAGAATCATCGCCTCATACGATCCAGAC KI4R | AACAAGGAAGAAAGAAGCATCGCCATCTACCTGGGCATTGGCCTCTGC E1371Q | CTCCTGTTTATTGTGCGGACTCTGCTGCTGCACCCAGCAATTTTCGGGTT GCATCATATTGGCATGCAGATGCGCATTGCTATGTTTTCCCTCATCTAC AAAAAGACACTGAAACTCAGCTCCCGGGTGCTGGACAAGATCTCCATC GGCCAACTGGTGTCTCTCCTGAGCAATAACTTGAATAAGTTCGACGAA GGGCTGGCCCTGGCACACTTCGTGTGGATTGCCCCCCTGCAGGTGGCCC TGCTGATGGGACTGATTTGGGAACTGCTGCAGGCTAGCGCTTTICTGCGG CCTGGGGTTCCTGATCGTGCTGGCACTGTTTCAGGCAGGCCTGGGCCGT ATGATGATGAAGTACAGAGACCAGAGGGCCGGGAAGATCTCCGAACG GCTCGTITATTACCTCTGAGATGATCGAGAACATTCAGTCTGTGAAAGCC TACTGCTGGGAGGAGGCTATGGAGAAGATGATCGAGAATCTGAGACAG ACCGAGCTGAAGCTGACCAGAAAGGCCGCCTACGTGAGGTACTTCAAC AGCAGTGCCTTCTTCTTCTCTGGCTTCTTCGTTGTGTITCTGAGCGTGCT GCCATACGCTCTCATCAAAGGCATCATCCTGCGGAAGATCTTCACCACC ATCAGCTTTTGCATCGTGCTTAGAATGGCCGTGACCCGGCAGITCCCAT GGGCCGTGCAAACTTGGTATGATTCCCTGGGCGCCATCAACAAAATCC AGGATTTCCTGCAGAAGCAGGAATACAAGACACTCGAATATAATCTCA CAACTACTGAGGTGGTTATGGAGAACGTGACTGCCTTCTGGGAGGAGG GGTTCGGAGAGCTTTTTGAGAAGGCAAAACAGAATAACAACAACCGCA AAACCAGCAACGGCGACGACAGCCTGTTCTICTCCAATTTTTCTCTCCT GGGAACACCCGTCCTCAAAGACATCAACTTTAAGATCGAGAGGGGACA GCTGCTCGCAGTCGCCGGATCCACAGGCGCCGGCAAGACCTCTCTGCT GATGGTTATCATGGGCGAACTGGAGCCATCCGAGGGCAAGATTAAGCA CAGTGGAAGAATCTCCTTTTGTAGCCAGTTCAGTTGGATTATGCCCGGC ACTATTAAGGAGAATATCATTTITGGGGTGAGCTATGATGAGTATCGGT ATCGGAGCGTTATCAAAGCCTGTCAGCTGGAGGAGGATATCAGCAAAT TCGCAGAGAAGGATAATATCGTGCTGGGGGAGGGGGGAATCACCCTGA GCGGAGGCCAGAGAGCCAGAATCTCACTGGCCCGGGCCGTCTACAAGG ACGCCGACCTTTACCTTCTGGACAGTCCCTTTGGATATCTGGATGTGCT GACTGAAAAGGAGATCTTCGAGTCTTGTGTGTGCAAGCTGATGGCTAA TAAGACCCGGATCCTAGTGACCAGTAAGATGGAGCACCTGAAGAAGGC AGACAAGATCTTGATTCTGCACGAGGGATCCTCTTACTTTTACGGCACC TTTAGCGAGCTGCAGAATCTCCAGCCCGATTTCTCATCTAAGCTGATGG GCTGTGATAGCTTCGACCAGTTCTCTGCCGAGCGCAGAAACAGCATCCT GACAGAGACACTGCACCGGTTTTCACTGGAGGGCGACGCCCCTGTCAG CTGGACCGAGACCAAAAAGCAGTCTTTCAAGCAGACAGGCGAGTTCGG CGAGAAGCGCAAAAACAGCATCCTGAATCCAATCAACTCTATAAGGAA GTTTAGCATCGTGCAGAAGACACCCCTCCAGATGAACGGCATCGAAGA GGACAGTGACGAGCCCCTGGAGCGGCGCCTGAGCCTCGTGCCTGACAG CGAACAGGGCGAGGCCATCCTGCCTAGGATCAGCGTGATTTCAACCGG GCCAACACTGCAGGCTAGGAGAAGACAGTCAGTGCTTAACCTGATGAC ACATAGCGTGAATCAGGGACAGAACATCCATCGAAAAACCACAGCCTC TACTCGCAAAGTGTCACTGGCTCCTCAGGCTAATCTGACAGAGCTGGA CATCTATAGCAGGAGGCTGAGCCAGGAGACAGGCCTGGAGATCAGTGA GGAGATCAACGAAGAGGACCTGAAGGAGTGCTTTTTCGATGACATGGA GAGTATCCCCGCCGTCACCACCTGGAATACCTACCTCCGGTACATCACA GTGCACAAGTCCCTCATCTTTGTGCTGATTTGGTGCCTCGTGATCTTTCT CGCAGAAGTGGCCGCCTCCCTGGTGGTGCTGTGGCTGTTGGGGAATAC TCCACTGCAGGACAAAGGCAATTCTACACACAGCAGGAATAATTCCTA TGCCGTGATTATCACCAGCACATCCTCTTACTACGTGTTCTACATCTAC GTGGGAGTGGCAGATACTCTGCTTGCAATGGGCTTCTTCAGGGGGCTG CCCCTGGTGCACACACTGATCACAGTGTCCAAGATCCTCCACCATAAA ATGCTCCACAGCGTGCTGCAGGCACCCATGAGCACCCTGAACACACTG AAGGCCGGCGGCATCCTGAATCGCTTTTCCAAAGACATCGCCATCCTCG ACGATCTCCTGCCACTGACCATCTTCGATTTTATCCAGCTGCTGCTGAT CGTGATCGGGGCCATCGCCGTGGTGGCCGTGCTGCAGCCATACATTTTC GTGGCTACAGTGCCCGTGATCGTTGCCTTTATCATGCTGAGAGCCTACT TCCTGCAGACTTCTCAGCAGCTGAAGCAGCTGGAGAGCGAAGGGAGAA GCCCCATCTTCACTCACCTGGTGACAAGCCTGAAGGGACTCTGGACCCT GAGAGCCTTCGGCCGGCAGCCCTATTTCGAGACCCTGTTTCACAAGGCC CTCAACCTGCACACAGCCAACTGGTTTCTCTACCTGTCCACCCTGAGGT NO: 50 Codon hCFTR- GGTTCCAGATGAGGATTGAAATGATCTTCGTGATTITITTCATCGCCGT GACATTCATTAGCATTCTGACCACCGGCGAGGGGGAGGGGAGAGTGGG CATCATCCTGACCCTTGCCATGAACATTATGTCCACACTGCAGTGGGCC GTGAATAGTTCAATCGACGTGGACAGTCTGATGAGGTCCGTGAGCCGG GTGTTCAAGTTCATTGACATGCCCACAGAGGGGAAACCCACCAAAAGC ACCAAGCCCTACAAGAACGGGCAGCTGTCCAAGGTTATGATCATCGAG AACTCTCACGTGAAGAAGGACGACATTTGGCCCAGCGGCGGCCAGATG ACAGTGAAAGATCTGACCGCCAAATACACCGAGGGAGGCAACGCCATC CTCGAAAACATTAGCTTCTCTATCAGCCCTGGACAGAGGGTGGGCCTG CTGGGCCGGACAGGCTCAGGGAAGAGTACTCTGCTGTCAGCATTCCTG AGGCTCCTGAACACAGAGGGCGAGATCCAGATTGACGGCGTGTCCTGG GACTCCATCACCCTGCAGCAGTGGCGGAAGGCTTTCGGGGTGATCCCC CAGAAGGTGTTCATCTTTAGCGGCACTTTCAGAAAGAATCTGGACCCTT ATGAGCAGTGGAGTGACCAGGAGATCTGGAAAGTGGCCGATGAGGTC GGACTGAGGAGCGTGATCGAGCAGTTTCCAGGGAAGCTGGACTITGTG CTGGTGGATGGCGGATGCGTGCTGTCTCACGGCCATAAACAGCTGATG TGTCTGGCCCGGTCCGTGCTGTCTAAGGCCAAGATCCTGCTGCTGGACC AACCCTCCGCCCACCTGGACCCCGTGACATACCAGATCATCAGGAGAA CTCTCAAGCAGGCCTTCGCCGACTGTACCGTGATTCTGTGCGAGCACCG CATTGAAGCTATGCTGGAGTGTCAGCAGTTCCTGGTGATCGAGGAAAA TAAGGTGAGGCAGTACGACAGCATCCAGAAGCTGCTGAACGAGCGCTC CCTGTTCCGCCAGGCTATCTCCCCATCAGACCGGGTGAAGCTCTTTCCC CACAGAAACTCCTCAAAGTGCAAGTCCAAGCCCCAGATCGCCGCCCTG AAGGAGGAGACCGAGGAGGAGGTGCAGGACACCAGGCTGTGA (SEQ 1D NO: 49) ATGCAGCGCTCGCCTCTGGAAAAGGCGAGCGTCGTGTCACGGCTATTC SEQID |TTTTCTTGGACCCGGCCCATTCTCAGGAAGGGCTACAGGCAGAGGCTG NO: 50 |GAGTTGAGCGACATCTATCAGATTCCTTCCGTGGACAGCGCCGACAAC CTGAGCGAGAAGCTGGAAAGGGAGTGGGACCGCGAACTGGCAAGCAA Codon | AAAGAACCCCAAGCTGATCAATGCCCTGAGAAGGTGTTTCTITTGGAG | Optimized —| ATTCATGTTCTACGGGATCTTTCTGTATCTGGGCGAGGTTACAAAGGCT hCFTR- | GTGCAGCCCCTGCTGCTCGGCAGAATCATCGCCTCATACGATCCAGAC 13EK14R | AACAAGGAAGAAAGAAGCATCGCCATCTACCTGGGCATTGGCCTCTGC CTCCTGTTTATTGTGCGGACTCTGCTGCTGCACCCAGCAATTTTCGGGTT GCATCATATTGGCATGCAGATGCGCATTGCTATGTTTTCCCTCATCTAC AAAAAGACACTGAAACTCAGCTCCCGGGTGCTGGACAAGATCTCCATC GGCCAACTGGTGTCTCTCCTGAGCAATAACTTGAATAAGTTCGACGAA GGGCTGGCCCTGGCACACTTCGTGTGGATTGCCCCCCTGCAGGTGGCCC TGCTGATGGGACTGATTTGGGAACTGCTGCAGGCTAGCGCTTTCTGCGG CCTGGGGTTCCTGATCGTGCTGGCACTGTTTCAGGCAGGCCTGGGCCGT ATGATGATGAAGTACAGAGACCAGAGGGCCGGGAAGATCTCCGAACG GCTCGTITATTACCTCTGAGATGATCGAGAACATTCAGTCTGTGAAAGCC TACTGCTGGGAGGAGGCTATGGAGAAGATGATCGAGAATCTGAGACAG ACCGAGCTGAAGCTGACCAGAAAGGCCGCCTACGTGAGGTACTTCAAC AGCAGIGCCTICTTCTICTCTGGCTTCTTCGTTGTGTTTCTGAGCGTGCT GCCATACGCTCTCATCAAAGGCATCATCCTGCGGAAGATCTTCACCACC ATCAGCTTTTGCATCGTGCTTAGAATGGCCGTGACCCGGCAGTTCCCAT GGGCCGTGCAAACTTGGTATGATTCCCTGGGCGCCATCAACAAAATCC AGGATTTCCTGCAGAAGCAGGAATACAAGACACTCGAATATAATCTCA CAACTACTGAGGTGGTTATGGAGAACGTGACTGCCTTCTGGGAGGAGG GGTTCGGAGAGCTTTTTGAGAAGGCAAAACAGAATAACAACAACCGCA AAACCGAGAACGGCGACGACAGCCTGTTCTTCTCCAATTTTTCTCTCCT GGGAACACCCGTCCTCAAAGACATCAACTTTAAGATCGAGAGGGGACA GCTGCTCGCAGTCGCCGGATCCACAGGCGCCGGCAAGACCTCTCTGCT GATGGTTATCATGGGCGAACTGGAGCCATCCGAGGGCAAGATTAAGCA CAGTGGAAGAATCTCCTTTTGTAGCCAGTTCAGTTGGATTATGCCCGGC ACTATTAAGGAGAATATCATTTTTGGGGTGAGCTATGATGAGTATCGGT ATCGGAGCGTTATCAAAGCCTGTCAGCTGGAGGAGGATATCAGCAAAT TCGCAGAGAAGGATAATATCGTGCTGGGGGAGGGGGGAATCACCCTGA GCGGAGGCCAGAGAGCCAGAATCTCACTGGCCCGGGCCGTCTACAAGG ACGCCGACCTTTACCTTCTGGACAGTCCCTTTGGATATCTGGATGTGCT GACTGAAAAGGAGATCTTCGAGTCTTGTGTGTGCAAGCTGATGGCTAA TAAGACCCGGATCCTAGTGACCAGTAAGATGGAGCACCTGAAGAAGGC AGACAAGATCTTGATTCTGCACGAGGGATCCTCTTACTTTTACGGCACC TTTAGCGAGCTGCAGAATCTCCAGCCCGATTTCTCATCTAAGCTGATGG GCTGTGATAGCTTCGACCAGTTCTCTGCCGAGCGCAGAAACGAAATCC TGACAGAGACACTGCACCGGTTTGAGCTGGAGGGCGACGCCCCTGTCA GCTGGACCGAGACCAAAAAGCAGGAATTCAAGCAGGAGGGCGAGTTC GGCGAGAAGCGCAAAAACGAAATCCTGAATCCAATCAACTCTATAAGG AAGTTTGAAATCGTGCAGAAGACACCCCTCCAGATGAACGGCATCGAA GAGGACAGTGACGAGCCCCTGGAGCGGCGCCTGAGCCTCGTGCCTGAC AGCGAACAGGGCGAGGCCATCCTGCCTAGGATCGAGGTGATTTCAACC GGGCCAACACTGCAGGCTAGGAGAAGACAGTCAGTGCTTAACCTGATG ACACATAGCGTGAATCAGGGACAGAACATCCATCGAAAAGAAGAAGC CGAAACTCGCAAAGTGGAGCTGGCTCCTCAGGCTAATCTGACAGAGCT GGACATCTATAGCAGGAGGCTGGAACAGGAGACAGGCCTGGAGATCA GTGAGGAGATCAACGAAGAGGACCTGAAGGAGTGCTTTTTCGATGACA TGGAGAGTATCCCCGCCGTCACCACCTGGAATACCTACCTCCGGTACAT CACAGTGCACAAGTCCCTCATCTTTGTGCTGATTTGGTGCCTCGTGATC TTTCTCGCAGAAGTGGCCGCCTCCCTGGTGGTGCTGTGGCTGTTGGGGA ATACTCCACTGCAGGACAAAGGCAATTCTACACACAGCAGGAATAATT CCTATGCCGTGATTATCACCAGCACATCCTCTTACTACGTGTTCTACAT CTACGTGGGAGTGGCAGATACTCTGCTTGCAATGGGCTTCTTCAGGGG GCTGCCCCTGGTGCACACACTGATCACAGTGTCCAAGATCCTCCACCAT AAAATGCTCCACAGCGTGCTGCAGGCACCCATGAGCACCCTGAACACA CTGAAGGCCGGCGGCATCCTGAATCGCTTTTCCAAAGACATCGCCATCC TCGACGATCTCCTGCCACTGACCATCTTCGATTTITATCCAGCTGCTGCT GATCGTGATCGGGGCCATCGCCGTGGTGGCCGTGCTGCAGCCATACAT TTTCGTGGCTACAGTGCCCGTGATCGTTGCCTTITATCATGCTGAGAGCC TACTTCCTGCAGACTTCTCAGCAGCTGAAGCAGCTGGAGAGCGAAGGG AGAAGCCCCATCTTCACTCACCTGGTGACAAGCCTGAAGGGACTCTGG Codon hCFTR- ACCCTGAGAGCCTTCGGCCGGCAGCCCTATTICGAGACCCTGITTCACA AGGCCCTCAACCTGCACACAGCCAACTGGTTTCTCTACCTGTCCACCCT GAGGTGGTTCCAGATGAGGATTGAAATGATCTTCGTGATTTTTITCATC GCCGTGACATTCATTAGCATTCTGACCACCGGCGAGGGGGAGGGGAGA GTGGGCATCATCCTGACCCTTGCCATGAACATTATGTCCACACTGCAGT GGGCCGTGAATAGTTCAATCGACGTGGACAGTCTGATGAGGTCCGTGA GCCGGGTGTTCAAGTTCATTGACATGCCCACAGAGGGGAAACCCACCA AAAGCACCAAGCCCTACAAGAACGGGCAGCTGTCCAAGGTTATGATCA TCGAGAACTCTCACGTGAAGAAGGACGACATTTGGCCCAGCGGCGGCC AGATGACAGTGAAAGATCTGACCGCCAAATACACCGAGGGAGGCAAC GCCATCCTCGAAAACATTAGCTTCTCTATCAGCCCTGGACAGAGGGTG GGCCTGCTGGGCCGGACAGGCTCAGGGAAGAGTACTCTGCTGTCAGCA TTCCTGAGGCTCCTGAACACAGAGGGCGAGATCCAGATTGACGGCGTG TCCTGGGACTCCATCACCCTGCAGCAGTGGCGGAAGGCTTTCGGGGTG ATCCCCCAGAAGGTGTTCATCTTTAGCGGCACTTTCAGAAAGAATCTGG ACCCTTATGAGCAGTGGAGTGACCAGGAGATCTGGAAAGTGGCCGATG AGGTCGGACTGAGGAGCGTGATCGAGCAGTTTCCAGGGAAGCTGGACT TTGTGCTGGTGGATGGCGGATGCGTGCTGTCTCACGGCCATAAACAGCT GATGTGTCTGGCCCGGTCCGTGCTGTCTAAGGCCAAGATCCTGCTGCTG GACGAACCCTCCGCCCACCTGGACCCCGTGACATACCAGATCATCAGG AGAACTCTCAAGCAGGCCTTCGCCGACTGTACCGTGATTCTGTGCGAGC ACCGCATTGAAGCTATGCTGGAGTGTCAGCAGTTCCTGGTGATCGAGG AAAATAAGGTGAGGCAGTACGACAGCATCCAGAAGCTGCTGAACGAG CGCTCCCTGTTCCGCCAGGCTATCTCCCCATCAGACCGGGTGAAGCTCT TTCCCCACAGAAACTCCTCAAAGTGCAAGTCCAAGCCCCAGATCGCCG CCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCAGGCTGTGA (SEQ ID NO: 50) ATGCAGCGCTCGCCTCTGGAAAAGGCGAGCGTCGTGTCACGGCTATIC SEQID |TTTTCTTGGACCCGGCCCATTCTCAGGAAGGGCTACAGGCAGAGGCTG NO: 51 |GAGTTGAGCGACATCTATCAGATTCCTTCCGTGGACAGCGCCGACAAC CTGAGCGAGAAGCTGGAAAGGGAGTGGGACCGCGAACTGGCAAGCAA Codon | AAAGAACCCCAAGCTGATCAATGCCCTGAGAAGGTGTTTCTITTGGAG Optimized | ATTCATGTTCTACGGGATCTTTCTGTATCTGGGCGAGGTTACAAAGGCT hCFTR- | GTGCAGCCCCTGCTGCTCGGCAGAATCATCGCCTCATACGATCCAGAC ISEK14R | AACAAGGAAGAAAGAAGCATCGCCATCTACCTGGGCATTGGCCTCTGC CTCCTGTTTATTGTGCGGACTCTGCTGCTGCACCCAGCAATTTTCGGGTT GCATCATATTGGCATGCAGATGCGCATTGCTATGTTTTCCCTCATCTAC AAAAAGACACTGAAACTCAGCTCCCGGGTGCTGGACAAGATCTCCATC GGCCAACTGGTGTCTCTCCTGAGCAATAACTTGAATAAGTTCGACGAA GGGCTGGCCCTGGCACACTTCGTGTGGATTGCCCCCCTGCAGGTGGCCC TGCTGATGGGACTGATTTGGGAACTGCTGCAGGCTAGCGCTTTCTGCGG CCTGGGGTTCCTGATCGTGCTGGCACTGTTTCAGGCAGGCCTGGGCCGT ATGATGATGAAGTACAGAGACCAGAGGGCCGGGAAGATCTCCGAACG GCTCGITATTACCTCTGAGATGATCGAGAACATTCAGTCTGTGAAAGCC TACTGCTGGGAGGAGGCTATGGAGAAGATGATCGAGAATCTGAGACAG ACCGAGCTGAAGCTGACCAGAAAGGCCGCCTACGTGAGGTACTTCAAC AGCAGTGCCTTCTTCTTCTCTGGCTTCTTCGTTGTGTTITCTGAGCGTGCT GCCATACGCTCTCATCAAAGGCATCATCCTGCGGAAGATCTTCACCACC ATCAGCTTTTGCATCGTGCTTAGAATGGCCGTGACCCGGCAGTTCCCAT GGGCCGTGCAAACTTGGTATGATTCCCTGGGCGCCATCAACAAAATCC AGGATTTCCTGCAGAAGCAGGAATACAAGACACTCGAATATAATCTCA CAACTACTGAGGTGGTTATGGAGAACGTGACTGCCTTCTGGGAGGAGG GGTTCGGAGAGCTTTTTGAGAAGGCAAAACAGAATAACAACAACCGCA AAACCGAGAACGGCGACGACAGCCTGTTCTTCTCCAATTTTTCTCTCCT GGGAACACCCGTCCTCAAAGACATCAACTTTAAGATCGAGAGGGGACA GCTGCTCGCAGTCGCCGGATCCACAGGCGCCGGCAAGACCTCTCTGCT GATGGTTATCATGGGCGAACTGGAGCCATCCGAGGGCAAGATTAAGCA CAGTGGAAGAATCTCCTTTTGTAGCCAGTTCAGTTGGATTATGCCCGGC ACTATTAAGGAGAATATCATTTTTGGGGTGAGCTATGATGAGTATCGGT ATCGGAGCGTTATCAAAGCCTGTCAGCTGGAGGAGGATATCAGCAAAT TCGCAGAGAAGGATAATATCGTGCTGGGGGAGGGGGGAATCACCCTGA GCGGAGGCCAGAGAGCCAGAATCTCACTGGCCCGGGCCGTCTACAAGG ACGCCGACCTTTACCTTCTGGACAGTCCCTTTGGATATCTGGATGTGCT GACTGAAAAGGAGATCTTCGAGTCTTGTGTGTGCAAGCTGATGGCTAA TAAGACCCGGATCCTAGTGACCAGTAAGATGGAGCACCTGAAGAAGGC AGACAAGATCTTGATTCTGCACGAGGGATCCTCTTACTTTTACGGCACC TTTAGCGAGCTGCAGAATCTCCAGCCCGATTTCTCATCTAAGCTGATGG GCTGTGATAGCTTCGACCAGTTCTCTGCCGAGCGCAGAAACGAAATCC TGACAGAGACACTGCACCGGTTTGAGCTGGAGGGCGACGCCCCTGTCA GCTGGACCGAGACCAAAAAGCAGGAATTCAAGCAGGAGGGCGAGTTC GGCGAGAAGCGCAAAAACGAAATCCTGAATCCAATCAACTCTATAAGG AAGTTTGAAATCGTGCAGAAGACACCCCTCCAGATGAACGGCATCGAA GAGGACAGTGACGAGCCCCTGGAGCGGCGCCTGGAACTCGTGCCTGAC AGCGAACAGGGCGAGGCCATCCTGCCTAGGATCGAGGTGATTTCAACC GGGCCAACACTGCAGGCTAGGAGAAGACAGGAAGTGCTTAACCTGATG ACACATAGCGTGAATCAGGGACAGAACATCCATCGAAAAGAAGAAGC CGAAACTCGCAAAGTGGAGCTGGCTCCTCAGGCTAATCTGACAGAGCT GGACATCTATAGCAGGAGGCTGGAACAGGAGACAGGCCTGGAGATCA GTGAGGAGATCAACGAAGAGGACCTGAAGGAGTGCTTTTTCGATGACA TGGAGAGTATCCCCGCCGTCACCACCTGGAATACCTACCTCCGGTACAT CACAGTGCACAAGTCCCTCATCTTTGTGCTGATTTGGTGCCTCGTGATC TTTCTCGCAGAAGTGGCCGCCTCCCTGGTGGTGCTGTGGCTGTTGGGGA ATACTCCACTGCAGGACAAAGGCAATTCTACACACAGCAGGAATAATT CCTATGCCGTGATTATCACCAGCACATCCTCTTACTACGTGTTCTACAT CTACGTGGGAGTGGCAGATACTCTGCTTGCAATGGGCTTCTTCAGGGG GCTGCCCCTGGTGCACACACTGATCACAGTGTCCAAGATCCTCCACCAT AAAATGCTCCACAGCGTGCTGCAGGCACCCATGAGCACCCTGAACACA CTGAAGGCCGGCGGCATCCTGAATCGCTTTTCCAAAGACATCGCCATCC TCGACGATCTCCTGCCACTGACCATCTTCGATTTITATCCAGCTGCTGCT GATCGTGATCGGGGCCATCGCCGTGGTGGCCGTGCTGCAGCCATACAT TTTCGTGGCTACAGTGCCCGTGATCGTTGCCTTTATCATGCTGAGAGCC Codon hCFTR- 13E TACTTCCTGCAGACTTCTCAGCAGCTGAAGCAGCTGGAGAGCGAAGGG AGAAGCCCCATCTTCACTCACCTGGTGACAAGCCTGAAGGGACTCTGG ACCCTGAGAGCCTTCGGCCGGCAGCCCTATTTCGAGACCCTGTTTCACA AGGCCCTCAACCTGCACACAGCCAACTGGTTTCTCTACCTGTCCACCCT GAGGTGGTTCCAGATGAGGATTGAAATGATCTTCGTGATTTTTTTCATC GCCGTGACATTCATTAGCATTCTGACCACCGGCGAGGGGGAGGGGAGA GTGGGCATCATCCTGACCCTTGCCATGAACATTATGTCCACACTGCAGT GGGCCGTGAATAGITCAATCGACGTGGACAGTCTGATGAGGTCCGTGA GCCGGGTGTTCAAGTTCATTGACATGCCCACAGAGGGGAAACCCACCA AAAGCACCAAGCCCTACAAGAACGGGCAGCTGTCCAAGGTTATGATCA TCGAGAACTCTCACGTGAAGAAGGACGACATTTGGCCCAGCGGCGGCC AGATGACAGTGAAAGATCTGACCGCCAAATACACCGAGGGAGGCAAC GCCATCCTCGAAAACATTAGCTTCTCTATCAGCCCTGGACAGAGGGTG GGCCTGCTGGGCCGGACAGGCTCAGGGAAGAGTACTCTGCTGTCAGCA TTCCTGAGGCTCCTGAACACAGAGGGCGAGATCCAGATTGACGGCGTG TCCTGGGACTCCATCACCCTGCAGCAGTGGCGGAAGGCTITCGGGGTG ATCCCCCAGAAGGTGITCATCTTTAGCGGCACTTTCAGAAAGAATCTGG ACCCTTATGAGCAGTGGAGTGACCAGGAGATCTGGAAAGTGGCCGATG AGGTCGGACTGAGGAGCGTGATCGAGCAGTTTCCAGGGAAGCTGGACT TTGTGCTGGTGGATGGCGGATGCGTGCTGTCTCACGGCCATAAACAGCT GATGTGTCTGGCCCGGTCCGTGCTGTCTAAGGCCAAGATCCTGCTGCTG GACGAACCCTCCGCCCACCTGGACCCCGTGACATACCAGATCATCAGG AGAACTCTCAAGCAGGCCTTCGCCGACTGTACCGTGATTCTGTGCGAGC ACCGCATTGAAGCTATGCTGGAGTGTCAGCAGTTCCTGGTGATCGAGG AAAATAAGGTGAGGCAGTACGACAGCATCCAGAAGCTGCTGAACGAG CGCTCCCTGTTCCGCCAGGCTATCTCCCCATCAGACCGGGTGAAGCTCT TTCCCCACAGAAACTCCTCAAAGTGCAAGTCCAAGCCCCAGATCGCCG CCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCAGGCTGTGA (SEQID NO: 51) ATGCAGCGCTCGCCTCTGGAAAAGGCGAGCGTCGTGTCAAAGCTATIC SEQID | TTITCTTGGACCCGGCCCATTCTCAGGAAGGGCTACAGGCAGAGGCTG NO: 52 |GAGTTGAGCGACATCTATCAGATTCCTTCCGTGGACAGCGCCGACAAC CTGAGCGAGAAGCTGGAAAGGGAGTGGGACCGCGAACTGGCAAGCAA Codon | AAAGAACCCCAAGCTGATCAATGCCCTGAGAAGGTGTTTCTITTGGAG Optimized- | ATTCATGTTCTACGGGATCTTTCTGTATCTGGGCGAGGTTACAAAGGCT hCFIR- | GTGCAGCCCCTGCTGCTCGGCAGAATCATCGCCTCATACGATCCAGAC BE | AACAAGGAAGAAAGAAGCATCGCCATCTACCTGGGCATTGGCCTCTGC CTCCTGTTTATTGTGCGGACTCTGCTGCTGCACCCAGCAATTTTCGGGTT GCATCATATTGGCATGCAGATGCGCATTGCTATGTTTTCCCTCATCTAC AAAAAGACACTGAAACTCAGCTCCCGGGTGCTGGACAAGATCTCCATC GGCCAACTGGTGTCTCTCCTGAGCAATAACTTGAATAAGTTCGACGAA GGGCTGGCCCTGGCACACTTCGTGTGGATTGCCCCCCTGCAGGTGGCCC TGCTGATGGGACTGATTTGGGAACTGCTGCAGGCTAGCGCTTTCTGCGG CCTGGGGTTCCTGATCGTGCTGGCACTGTTTCAGGCAGGCCTGGGCCGT ATGATGATGAAGTACAGAGACCAGAGGGCCGGGAAGATCTCCGAACG GCTCGTTATTACCTCTGAGATGATCGAGAACATTCAGTCTGTGAAAGCC TACTGCTGGGAGGAGGCTATGGAGAAGATGATCGAGAATCTGAGACAG ACCGAGCTGAAGCTGACCAGAAAGGCCGCCTACGTGAGGTACTTCAAC AGCAGTGCCTTCTTCTTCTCTGGCTTCTTCGTTGTGTTITCTGAGCGTGCT GCCATACGCTCTCATCAAAGGCATCATCCTGCGGAAGATCTTCACCACC ATCAGCTTTTGCATCGTGCTTAGAATGGCCGTGACCCGGCAGTTCCCAT GGGCCGTGCAAACTTGGTATGATTCCCTGGGCGCCATCAACAAAATCC AGGATTTCCTGCAGAAGCAGGAATACAAGACACTCGAATATAATCTCA CAACTACTGAGGTGGTTATGGAGAACGTGACTGCCTTCTGGGAGGAGG GGTTCGGAGAGCTTTTTGAGAAGGCAAAACAGAATAACAACAACCGCA AAACCGAGAACGGCGACGACAGCCTGTTCTTCTCCAATTTTTCTCTCCT GGGAACACCCGTCCTCAAAGACATCAACTTTAAGATCGAGAGGGGACA GCTGCTCGCAGTCGCCGGATCCACAGGCGCCGGCAAGACCTCTCTGCT GATGGTTATCATGGGCGAACTGGAGCCATCCGAGGGCAAGATTAAGCA CAGTGGAAGAATCTCCTTTTGTAGCCAGTTCAGTTGGATTATGCCCGGC ACTATTAAGGAGAATATCATTTTTGGGGTGAGCTATGATGAGTATCGGT ATCGGAGCGTTATCAAAGCCTGTCAGCTGGAGGAGGATATCAGCAAAT TCGCAGAGAAGGATAATATCGTGCTGGGGGAGGGGGGAATCACCCTGA GCGGAGGCCAGAGAGCCAGAATCTCACTGGCCCGGGCCGTCTACAAGG ACGCCGACCTTTACCTTCTGGACAGTCCCTTTGGATATCTGGATGTGCT GACTGAAAAGGAGATCTTCGAGTCTTGTGTGTGCAAGCTGATGGCTAA TAAGACCCGGATCCTAGTGACCAGTAAGATGGAGCACCTGAAGAAGGC AGACAAGATCTTGATTCTGCACGAGGGATCCTCTTACTTTTACGGCACC TTTAGCGAGCTGCAGAATCTCCAGCCCGATTTCTCATCTAAGCTGATGG GCTGTGATAGCTTCGACCAGTTCTCTGCCGAGCGCAGAAACGAAATCC TGACAGAGACACTGCACCGGTTTGAGCTGGAGGGCGACGCCCCTGTCA GCTGGACCGAGACCAAAAAGCAGGAATTCAAGCAGGAGGGCGAGTTC GGCGAGAAGCGCAAAAACGAAATCCTGAATCCAATCAACTCTATAAGG AAGTTTGAAATCGTGCAGAAGACACCCCTCCAGATGAACGGCATCGAA GAGGACAGTGACGAGCCCCTGGAGCGGCGCCTGAGCCTCGTGCCTGAC AGCGAACAGGGCGAGGCCATCCTGCCTAGGATCGAGGTGATTTCAACC GGGCCAACACTGCAGGCTAGGAGAAGACAGTCAGTGCTTAACCTGATG ACACATAGCGTGAATCAGGGACAGAACATCCATCGAAAAGAAGAAGC CGAAACTCGCAAAGTGGAGCTGGCTCCTCAGGCTAATCTGACAGAGCT GGACATCTATAGCAGGAGGCTGGAACAGGAGACAGGCCTGGAGATCA GTGAGGAGATCAACGAAGAGGACCTGAAGGAGTGCTTTTTCGATGACA TGGAGAGTATCCCCGCCGTCACCACCTGGAATACCTACCTCCGGTACAT CACAGTGCACAAGTCCCTCATCTTTGTGCTGATTTGGTGCCTCGTGATC TTTCTCGCAGAAGTGGCCGCCTCCCTGGTGGTGCTGTGGCTGTTGGGGA ATACTCCACTGCAGGACAAAGGCAATTCTACACACAGCAGGAATAATT CCTATGCCGTGATTATCACCAGCACATCCTCTTACTACGTGTTCTACAT CTACGTGGGAGTGGCAGATACTCTGCTTGCAATGGGCTTCTTCAGGGG GCTGCCCCTGGTGCACACACTGATCACAGTGTCCAAGATCCTCCACCAT AAAATGCTCCACAGCGTGCTGCAGGCACCCATGAGCACCCTGAACACA CTGAAGGCCGGCGGCATCCTGAATCGCTTTTCCAAAGACATCGCCATCC TCGACGATCTCCTGCCACTGACCATCTTCGATTITATCCAGCTGCTGCT GATCGTGATCGGGGCCATCGCCGTGGTGGCCGTGCTGCAGCCATACAT TTTCGTGGCTACAGTGCCCGTGATCGTTGCCTTTATCATGCTGAGAGCC TACTTCCTGCAGACTTCTCAGCAGCTGAAGCAGCTGGAGAGCGAAGGG AGAAGCCCCATCTTCACTCACCTGGTGACAAGCCTGAAGGGACTCTGG ACCCTGAGAGCCTTCGGCCGGCAGCCCTATTTCGAGACCCTGTTTCACA AGGCCCTCAACCTGCACACAGCCAACTGGTTTCTCTACCTGTCCACCCT GAGGTGGTTCCAGATGAGGATTGAAATGATCTICGTGATTTTTTTCATC GCCGTGACATTCATTAGCATTCTGACCACCGGCGAGGGGGAGGGGAGA GTGGGCATCATCCTGACCCTTGCCATGAACATTATGTCCACACTGCAGT GGGCCGTGAATAGTTCAATCGACGTGGACAGTCTGATGAGGTCCGTGA GCCGGGTGTTCAAGTTCATTGACATGCCCACAGAGGGGAAACCCACCA AAAGCACCAAGCCCTACAAGAACGGGCAGCTGTCCAAGGTTATGATCA TCGAGAACTCTCACGTGAAGAAGGACGACATTITGGCCCAGCGGCGGCC AGATGACAGTGAAAGATCTGACCGCCAAATACACCGAGGGAGGCAAC GCCATCCTCGAAAACATTAGCTTCTCTATCAGCCCTGGACAGAGGGTG GGCCTGCTGGGCCGGACAGGCTCAGGGAAGAGTACTCTGCTGTCAGCA TTCCTGAGGCTCCTGAACACAGAGGGCGAGATCCAGATTGACGGCGTG TCCTGGGACTCCATCACCCTGCAGCAGTGGCGGAAGGCTTTCGGGGTG ATCCCCCAGAAGGTGTTCATCTTTAGCGGCACTTTCAGAAAGAATCTGG ACCCTTATGAGCAGTGGAGTGACCAGGAGATCTGGAAAGTGGCCGATG AGGTCGGACTGAGGAGCGTGATCGAGCAGTTTCCAGGGAAGCTGGACT TTGTGCTGGTGGATGGCGGATGCGTGCTGTCTCACGGCCATAAACAGCT GATGTGTCTGGCCCGGTCCGTGCTGTCTAAGGCCAAGATCCTGCTGCTG GACGAACCCTCCGCCCACCTGGACCCCGTGACATACCAGATCATCAGG AGAACTCTCAAGCAGGCCTTCGCCGACTGTACCGTGATTCTGTGCGAGC ACCGCATTGAAGCTATGCTGGAGTGTCAGCAGTTCCTGGTGATCGAGG AAAATAAGGTGAGGCAGTACGACAGCATCCAGAAGCTGCTGAACGAG CGCTCCCTGTTCCGCCAGGCTATCTCCCCATCAGACCGGGTGAAGCTCT TTCCCCACAGAAACTCCTCAAAGTGCAAGTCCAAGCCCCAGATCGCCG CCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCAGGCTGTGA |sEQ ID NO: 52)

[0125] Tn some embodiments, a suitable mRNA sequence may be an mRNA sequence encoding a homolog or an analog of human CFTR (hCFTR) protein. For example, a homolog or an analog of hCFTR protein may be a modified hCFTR protein containing one or more amino acid substitutions, deletions, and / or insertions as compared to a wild-type or naturally-occurring hCFTR protein while retaining substantial hCFTR protein activity. Tn some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more homologous to SEQ ID NO: 3, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, or SEQ ID NO: 44. In some embodiments, an mRNA suitable for the present invention encodes a protein substantially identical to an engineered hCFTR protein. In some embodiments, an mRNA suitable for the present invention encodes an amino acid sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, or SEQ ID NO: 44. In some embodiments, an mRNA suitable for the present invention encodes a fragment or a portion of an engineered hCFTR protein. In some embodiments, an mRNA suitable for the present invention encodes a fragment or a portion of an engineered hCFTR protein, wherein the fragment or portion of the protein still maintains CFTR activity similar to that of the full-length engineered protein, In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, or SEQ ID NO: 48.

[0126] In some embodiments, an mRNA suitable for the present invention has a nucleotide sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to any one of SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39 or SEQ ID NO: 40.

[0127] In some embodiments, a suitable mRNA encodes a fusion protein comprising a full length, fragment or portion of an engineered hCFTR protein fused to another protein (e.g., an N or C terminal fusion). In some embodiments, the protein fused to the mRNA encoding a full length, fragment or portion of an engineered hCFTR protein encodes a signal or a cellular targeting sequence. ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTITIG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTITCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTITGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTICTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 21) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCTGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TITGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TITTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATTTCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTITCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTICTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTTG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTICTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTIGTIGTIGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTITCATCTTICTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTTGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 22) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTITTITTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTITGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTITTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTITTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTITGGAGTGTICCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTITG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTTGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACACACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTITGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTIGCACACTGCCAACTGGTITCTITTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTICTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTTGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTITGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTITCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 23) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTITTIGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTICACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTITIG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCAACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTIGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 24) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TITGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TITTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTICTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTTG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCTATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTTGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TICTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTITGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTITTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTITCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 25) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTITTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATTTCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTICACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTTG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTITGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCAACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTIGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 26) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCTGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TITGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTIGGGCTGGGCT TITTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTICACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCCGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTITCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTITG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTICTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTTGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTICTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAGCCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTITCATCTTICTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTITTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 27) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TITTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTICACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTTG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCTATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTITAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTTGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TICTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACACACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTICTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTICAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 28) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTITTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTITCTGGAGGTTCATGTTITTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTTGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTITTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGICCTA TGATGAGTACCGCTACCGGTCCGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTTG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAGTCTGAGGGCCGGAGCCCCATTITTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTITGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 29) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTTTTTTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCTGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTITCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTITATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTIGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TITTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATTTCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTICTTCTTCAGTGGCTTCTTIGTTGTICTTCCTGTICTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTTTTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTTTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTTGGAGTGTCCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTTITG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TICTCAATCCTATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACACACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTTTGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTITGAGACACTGTTCCACAAG GCCCTGAACTTGCACACTGCCAACTGGTTTCTTTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTCTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTTTGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTTTCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 30) ATGCAGAGAAGCCCCCTGGAGAAGGCCTCTGTGGTGAGCAAGCTGTTCTTCAGCTG GACCAGACCCATCCTGAGAAAGGGCTACAGACAGAGACTGGAGCTGTICTGACATCT ACCAGATCCCCTCTGTGGACTCTGCCGACAACCTGTCTGAGAAGCTGGAGAGAGAG TGGGACAGAGAGCTGGCCAGCAAGAAGAACCCCAAGCTGATCAATGCCCTGAGAA GATGCTTCTTCTGGAGATTCATGTTCTATGGCATCTTCCTGTACCTGGGAGAGGTGAC CAAGGCCGTGCAGCCCCTGCTGCTGGGCAGGATCATTGCCAGCTATGACCCTGACA ACAAGGAGGAGAGAAGCATTGCCATCTACCTGGGCATTGGCCTGTGCCTGCTGTTCA TTGTGAGAACCCTGCTGCTGCACCCTGCCATCTTTGGCCTGCACCACATTGGCATGC AGATGAGAATTGCCATGTTCAGCCTGATCTACAAGAAGACCCTGAAGCTGAGCAGC AGAGTGCTGGACAAGATCAGCATTGGCCAGCTGGTGAGCCTGCTGAGCAACAACCT GAACAAGTTTGATGAGGGCCTGGCCCTGGCCCACTTTGTGTGGATTGCCCCCCTGCA GGTGGCCCTGCTGATGGGCCTGATCTGGGAGCTGCTGCAGGCCTCTGCCTTCTGTGG CCTGGGCTTCCTGATTGTGCTGGCCCTGTTCCAGGCCGGCCTGGGCAGAATGATGAT GAAGTACAGAGACCAGAGAGCCGGCAAGATCTCTGAGAGACTGGTGATCACCTCTG AGATGATTGAGAACATCCAGTCTGTGAAGGCCTACTGCTGGGAGGAGGCCATGGAG AAGATGATTGAGAACCTGAGACAGACAGAGCTGAAGCTGACCAGGAAGGCCGCCTA TGTGAGATACTTCAACAGCTCTGCCTTCTTCTTCTCTGGCTTCTTTGTGGTGTTCCTGT CTGTGCTGCCCTATGCCCTGATCAAGGGCATCATCCTGAGGAAGATCTTCACCACCA TCAGCTTCTGCATTGTGCTGAGGATGGCCGTGACCAGGCAGTTCCCCTGGGCCGTGC AGACCTGGTATGACAGCCTGGGGGCCATCAACAAGATCCAGGACTTCCTGCAGAAG CAGGAGTACAAGACCCTGGAGTACAACCTGACCACCACAGAGGTGGTGATGGAGAA TGTGACAGCCTTCTGGGAGGAGGGCTTTGGAGAGCTGTTTGAGAAGGCCAAGCAGA ACAACAACAACAGAAAGACCAGCAATGGAGATGACAGCCTGTTCTTCAGCAACTTC AGCCTGCTGGGCACCCCTGTGCTGAAGGACATCAACTTCAAGATTGAGAGGGGCCA GCTGCTGGCCGTGGCCGGCAGCACAGGAGCCGGCAAGACCAGCCTGCTGATGGTGA TCATGGGAGAGCTGGAGCCCTCTGAGGGCAAGATCAAGCACTCTGGCAGAATCAGC TTCTGCAGCCAGTTCAGCTGGATCATGCCTGGCACCATCAAGGAGAACATCATCTTT GGGGTGAGCTATGATGAGTACAGGTACAGATCTGTGATCAAGGCCTGCCAGCTGGA GGAGGACATCTCCAAGTTTGCCGAGAAGGACAACATTGTGCTGGGGGAGGGAGGCA TCACCCTGTCTGGGGGCCAGAGAGCCAGAATCAGCCTGGCCAGAGCCGTGTACAAG GATGCCGACCTGTACCTGCTGGACAGCCCCTTTGGCTACCTGGATGTGCTGACAGAG AAGGAGATCTTTGAGAGCTGTGTGTGCAAGCTGATGGCCAACAAGACCAGGATCCT GGTGACCAGCAAGATGGAGCACCTGAAGAAGGCCGACAAGATCCTGATCCTGCATG AGGGCAGCAGCTACTTCTATGGCACCTTCTCTGAGCTGCAGAACCTGCAGCCTGACT TCAGCAGCAAGCTGATGGGCTGTGACAGCTTTGACCAGTTCTCTGCTGAGAGAAGA AACAGCATCCTGACAGAGACCCTGCACAGGTTCAGCCTGGAGGGGGATGCCCCTGT GAGCTGGACAGAGACCAAGAAGCAGAGCTTCAAGCAGACAGGAGAGTTTGGGGAG AAGAGGAAGAACAGCATCCTGAACCCCATCAACAGCATCAGGAAGTTCAGCATTGT GCAGAAGACCCCCCTGCAGATGAATGGCATTGAGGAGGACTCTGATGAGCCCCTGG AGAGAAGACTGAGCCTGGTGCCAGACTCTGAGCAGGGAGAGGCCATCCTGCCCAGG ATCTCTGTGATCAGCACAGGCCCCACCCTGCAGGCCAGAAGAAGACAGTCTGTGCT GAACCTGATGACCCACTCTGTGAACCAGGGCCAGAATATCCACAGAAAGACCACAG CCAGCACCAGAAAGGTGAGCCTGGCCCCCCAGGCCAACCTGACAGAGCTGGACATC TACAGCAGAAGGCTGAGCCAGGAGACAGGCCTGGAGATCTCTGAGGAGATCAATGA GGAGGACCTGAAGGAGTGCTTCTTTGATGACATGGAGAGCATCCCTGCCGTGACCA CCTGGAACACCTACCTGAGATACATCACAGTGCACAAGAGCCTGATCTTTGTGCTGA TCTGGTGCCTGGTGATCTTCCTGGCCGAGGTGGCCGCCAGCCTGGTGGTGCTGTGGC TGCTGGGCAACACCCCCCTGCAGGACAAGGGCAACAGCACCCACAGCAGAAACAAC AGCTATGCTGTGATCATCACCAGCACCAGCAGCTACTATGTGTTCTACATCTATGTG GGAGTGGCTGACACCCTGCTGGCCATGGGCTTCTTCAGAGGCCTGCCCCTGGTGCAC ACCCTGATCACAGTGAGCAAGATCCTGCACCACAAGATGCTGCACTCTGTGCTGCAG GCCCCCATGAGCACCCTGAACACCCTGAAGGCTGGAGGCATCCTGAACAGATTCAG CAAGGACATTGCCATCCTGGATGACCTGCTGCCCCTGACCATCTTTGACTTCATCCA GCTGCTGCTGATTGTGATTGGAGCCATTGCCGTGGTGGCCGTGCTGCAGCCCTACAT CTTTGTGGCCACAGTGCCTGTGATTGTGGCCTTCATCATGCTGAGGGCCTACTTCCTG CAGACCAGCCAGCAGCTGAAGCAGCTGGAGTCTGAGGGCAGAAGCCCCATCTTCAC CCACCTGGTGACCAGCCTGAAGGGCCTGTGGACCCTGAGGGCCTTTGGCAGACAGC CCTACTTTGAGACCCTGTTCCACAAGGCCCTGAACCTGCACACAGCCAACTGGTTCC TGTACCTGAGCACCCTGAGATGGTTCCAGATGAGGATTGAGATGATCTTTGTGATCT TCTTCATTGCCGTGACCTTCATCAGCATCCTGACCACAGGGGAGGGCGAGGGCAGA GTGGGCATCATCCTGACCCTGGCCATGAACATCATGAGCACCCTGCAGTGGGCCGTG AACAGCAGCATTGATGTGGACAGCCTGATGAGATCTGTGAGCAGAGTGTTCAAGTT CATTGACATGCCCACAGAGGGCAAGCCCACCAAGAGCACCAAGCCCTACAAGAATG GCCAGCTGAGCAAGGTGATGATCATTGAGAACAGCCATGTGAAGAAGGATGACATC TGGCCCTCTGGAGGCCAGATGACAGTGAAGGACCTGACAGCCAAGTACACAGAGGG GGGCAATGCCATCCTGGAGAACATCAGCTTCAGCATCAGCCCTGGCCAGAGGGTGG GCCTGCTGGGCAGAACAGGCTCTGGCAAGAGCACCCTGCTGTCTGCCTTCCTGAGGC TGCTGAACACAGAGGGAGAGATCCAGATTGATGGGGTGAGCTGGGACAGCATCACC CTGCAGCAGTGGAGGAAGGCCTTTGGGGTGATCCCCCAGAAGGTGTTCATCTTCTCT GGCACCTTCAGGAAGAACCTGGACCCCTATGAGCAGTGGTCTGACCAGGAGATCTG GAAGGTGGCCGATGAGGTGGGCCTGAGATCTGTGATTGAGCAGTTCCCTGGCAAGC TGGACTTTGTGCTGGTGGATGGAGGCTGTGTGCTGAGCCATGGCCACAAGCAGCTGA TGTGCCTGGCCAGATCTGTGCTGAGCAAGGCCAAGATCCTGCTGCTGGATGAGCCCT CTGCCCACCTGGACCCTGTGACCTACCAGATCATCAGAAGAACCCTGAAGCAGGCC TTTGCCGACTGCACAGTGATCCTGTGTGAGCACAGAATTGAGGCCATGCTGGAGTGC CAGCAGTTCCTGGTGATTGAGGAGAACAAGGTGAGGCAGTATGACAGCATCCAGAA GCTGCTGAATGAGAGAAGCCTGTTCAGACAGGCCATCAGCCCCTCTGACAGAGTGA AGCTGTTCCCCCACAGGAACAGCAGCAAGTGCAAGAGCAAGCCCCAGATTGCCGCC CTGAAGGAGGAGACAGAGGAGGAGGTGCAGGACACCAGACTGTGA (SEQ ID NO: 31) ATGCAGAGGAGCCCCCTGGAGAAGGCCAGCGTGGTGAGCAAGCTGTTCTTCAGCTG GACCAGGCCCATCCTGAGGAAGGGCTACAGGCAGAGGCTGGAGCTGAGCGACATCT ACCAGATCCCCAGCGTGGACAGCGCCGACAACCTGAGCGAGAAGCTGGAGAGGGA GTGGGACAGGGAGCTGGCCAGCAAGAAGAACCCCAAGCTGATCAACGCCCTGAGG AGGTGCTTCTTCTGGAGGTTCATGTTCTACGGCATCTTCCTGTACCTGGGCGAGGTG ACCAAGGCCGTGCAGCCCCTGCTGCTGGGCAGGATCATCGCCAGCTACGACCCCGA CAACAAGGAGGAGAGGAGCATCGCCATCTACCTGGGCATCGGCCTGTGCCTGCTGT TCATCGTGAGGACCCTGCTGCTGCACCCCGCCATCTTCGGCCTGCACCACATCGGCA TGCAGATGAGGATCGCCATGTTCAGCCTGATCTACAAGAAGACCCTGAAGCTGAGC AGCAGGGTGCTGGACAAGATCAGCATCGGCCAGCTGGTGAGCCTGCTGAGCAACAA CCTGAACAAGTTCGACGAGGGCCTGGCCCTGGCCCACTTCGTGTGGATCGCCCCCCT GCAGGTGGCCCTGCTGATGGGCCTGATCTGGGAGCTGCTGCAGGCCAGCGCCTTCTG CGGCCTGGGCTTCCTGATCGTGCTGGCCCTGTTCCAGGCCGGCCTGGGCAGGATGAT GATGAAGTACAGGGACCAGAGGGCCGGCAAGATCAGCGAGAGGCTGGTGATCACC AGCGAGATGATCGAGAACATCCAGAGCGTGAAGGCCTACTGCTGGGAGGAGGCCAT GGAGAAGATGATCGAGAACCTGAGGCAGACCGAGCTGAAGCTGACCAGGAAGGCC GCCTACGTGAGGTACTTCAACAGCAGCGCCTTCTTCTTCAGCGGCTTCTTCGTGGTGT TCCTGAGCGTGCTGCCCTACGCCCTGATCAAGGGCATCATCCTGAGGAAGATCTTCA CCACCATCAGCTTCTGCATCGTGCTGAGGATGGCCGTGACCAGGCAGTTCCCCTGGG CCGTGCAGACCTGGTACGACAGCCTGGGCGCCATCAACAAGATCCAGGACTTCCTG CAGAAGCAGGAGTACAAGACCCTGGAGTACAACCTGACCACCACCGAGGTGGTGAT GGAGAACGTGACCGCCTTCTGGGAGGAGGGCTTCGGCGAGCTGTTCGAGAAGGCCA AGCAGAACAACAACAACAGGAAGACCAGCAACGGCGACGACAGCCTGTTCTTCAGC AACTTCAGCCTGCTGGGCACCCCCGTGCTGAAGGACATCAACTTCAAGATCGAGAG GGGCCAGCTGCTGGCCGTGGCCGGCAGCACCGGCGCCGGCAAGACCAGCCTGCTGA TGGTGATCATGGGCGAGCTGGAGCCCAGCGAGGGCAAGATCAAGCACAGCGGCAG GATCAGCTTCTGCAGCCAGTTCAGCTGGATCATGCCCGGCACCATCAAGGAGAACA TCATCTTCGGCGTGAGCTACGACGAGTACAGGTACAGGAGCGTGATCAAGGCCTGC CAGCTGGAGGAGGACATCAGCAAGTTCGCCGAGAAGGACAACATCGTGCTGGGCGA GGGCGGCATCACCCTGAGCGGCGGCCAGAGGGCCAGGATCAGCCTGGCCAGGGCCG TGTACAAGGACGCCGACCTGTACCTGCTGGACAGCCCCTTCGGCTACCTGGACGTGC TGACCGAGAAGGAGATCTTCGAGAGCTGCGTGTGCAAGCTGATGGCCAACAAGACC AGGATCCTGGTGACCAGCAAGATGGAGCACCTGAAGAAGGCCGACAAGATCCTGAT CCTGCACGAGGGCAGCAGCTACTTCTACGGCACCTTCAGCGAGCTGCAGAACCTGC AGCCCGACTTCAGCAGCAAGCTGATGGGCTGCGACAGCTTCGACCAGTTCAGCGCC GAGAGGAGGAACAGCATCCTGACCGAGACCCTGCACAGGTTCAGCCTGGAGGGCGA CGCCCCCGTGAGCTGGACCGAGACCAAGAAGCAGAGCTTCAAGCAGACCGGCGAGT TCGGCGAGAAGAGGAAGAACAGCATCCTGAACCCCATCAACAGCATCAGGAAGTTC AGCATCGTGCAGAAGACCCCCCTGCAGATGAACGGCATCGAGGAGGACAGCGACG AGCCCCTGGAGAGGAGGCTGAGCCTGGTGCCCGACAGCGAGCAGGGCGAGGCCATC CTGCCCAGGATCAGCGTGATCAGCACCGGCCCCACCCTGCAGGCCAGGAGGAGGCA GAGCGTGCTGAACCTGATGACCCACAGCGTGAACCAGGGCCAGAACATCCACAGGA AGACCACCGCCAGCACCAGGAAGGTGAGCCTGGCCCCCCAGGCCAACCTGACCGAG CTGGACATCTACAGCAGGAGGCTGAGCCAGGAGACCGGCCTGGAGATCAGCGAGG AGATCAACGAGGAGGACCTGAAGGAGTGCTTCTTCGACGACATGGAGAGCATCCCC GCCGTGACCACCTGGAACACCTACCTGAGGTACATCACCGTGCACAAGAGCCTGAT CTTCGTGCTGATCTGGTGCCTGGTGATCTTCCTGGCCGAGGTGGCCGCCAGCCTGGT GGTGCTGTGGCTGCTGGGCAACACCCCCCTGCAGGACAAGGGCAACAGCACCCACA GCAGGAACAACAGCTACGCCGTGATCATCACCAGCACCAGCAGCTACTACGTGTTC TACATCTACGTGGGCGTGGCCGACACCCTGCTGGCCATGGGCTTCTTCAGGGGCCTG CCCCTGGTGCACACCCTGATCACCGTGAGCAAGATCCTGCACCACAAGATGCTGCAC AGCGTGCTGCAGGCCCCCATGAGCACCCTGAACACCCTGAAGGCCGGCGGCATCCT GAACAGGTTCAGCAAGGACATCGCCATCCTGGACGACCTGCTGCCCCTGACCATCTT CGACTTCATCCAGCTGCTGCTGATCGTGATCGGCGCCATCGCCGTGGTGGCCGTGCT GCAGCCCTACATCTTCGTGGCCACCGTGCCCGTGATCGTGGCCTTCATCATGCTGAG GGCCTACTTCCTGCAGACCAGCCAGCAGCTGAAGCAGCTGGAGAGCGAGGGCAGGA GCCCCATCTTCACCCACCTGGTGACCAGCCTGAAGGGCCTGTGGACCCTGAGGGCCT TCGGCAGGCAGCCCTACTTCGAGACCCTGTTCCACAAGGCCCTGAACCTGCACACCG CCAACTGGTTCCTGTACCTGAGCACCCTGAGGTGGTTCCAGATGAGGATCGAGATGA TCTTCGTGATCTTCTTCATCGCCGTGACCTTCATCAGCATCCTGACCACCGGCGAGG GCGAGGGCAGGGTGGGCATCATCCTGACCCTGGCCATGAACATCATGAGCACCCTG CAGTGGGCCGTGAACAGCAGCATCGACGTGGACAGCCTGATGAGGAGCGTGAGCAG GGTGTTCAAGTTCATCGACATGCCCACCGAGGGCAAGCCCACCAAGAGCACCAAGC CCTACAAGAACGGCCAGCTGAGCAAGGTGATGATCATCGAGAACAGCCACGTGAAG AAGGACGACATCTGGCCCAGCGGCGGCCAGATGACCGTGAAGGACCTGACCGCCAA GTACACCGAGGGCGGCAACGCCATCCTGGAGAACATCAGCTTCAGCATCAGCCCCG GCCAGAGGGTGGGCCTGCTGGGCAGGACCGGCAGCGGCAAGAGCACCCTGCTGAGC GCCTTCCTGAGGCTGCTGAACACCGAGGGCGAGATCCAGATCGACGGCGTGAGCTG GGACAGCATCACCCTGCAGCAGTGGAGGAAGGCCTTCGGCGTGATCCCCCAGAAGG TGTTCATCTTCAGCGGCACCTTCAGGAAGAACCTGGACCCCTACGAGCAGTGGAGC GACCAGGAGATCTGGAAGGTGGCCGACGAGGTGGGCCTGAGGAGCGTGATCGAGC AGTTCCCCGGCAAGCTGGACTTCGTGCTGGTGGACGGCGGCTGCGTGCTGAGCCAC GGCCACAAGCAGCTGATGTGCCTGGCCAGGAGCGTGCTGAGCAAGGCCAAGATCCT GCTGCTGGACGAGCCCAGCGCCCACCTGGACCCCGTGACCTACCAGATCATCAGGA GGACCCTGAAGCAGGCCTTCGCCGACTGCACCGTGATCCTGTGCGAGCACAGGATC GAGGCCATGCTGGAGTGCCAGCAGTTCCTGGTGATCGAGGAGAACAAGGTGAGGCA GTACGACAGCATCCAGAAGCTGCTGAACGAGAGGAGCCTGTTCAGGCAGGCCATCA GCCCCAGCGACAGGGTGAAGCTGTTCCCCCACAGGAACAGCAGCAAGTGCAAGAGC AAGCCCCAGATCGCCGCCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCA GGCTGTGA (SEQ ID NO: 32) ATGCAGAGATCCCCTCTGGAGAAGGCCTCAGTGGTGTCCAAGCTTTTCTICTCCTGG ACCAGGCCCATTTTAAGAAAGGGCTACAGGCAGAGACTTGAGCTGTCTGACATCTAT CAGATCCCTTCTGTGGATTCTGCTGACAATCTTAGTGAAAAATTGGAAAGGGAGTGG GACAGAGAGCTGGCAAGTAAAAAGAACCCCAAGCTGATTAATGCCCTGAGGCGCTG CTTTTTTTGGAGATTCATGTTCTATGGCATATTCCTCTACCTTGGAGAAGTAACCAAA GCTGTACAGCCTCTCCTCCTTGGCAGAATCATTGCCTCCTATGATCCTGATAACAAG GAGGAGAGAAGCATAGCCATCTACCTGGGCATTGGGCTGTGCCTCTTGTTTATTGTG AGGACCCTTCTCTTGCACCCTGCCATCTTTGGCCTTCATCACATTGGCATGCAAATGA GAATAGCAATGTTTAGTCTTATTTACAAAAAAACATTAAAACTCTCTTCCAGGGTGT TGGACAAGATCAGTATTGGACAACTGGTCAGCCTGCTGAGCAACAACCTGAACAAG TTTGATGAAGGACTGGCCCTGGCCCACTTITGTCTGGATTGCCCCCCTTCAGGTGGCTC TITTGATGGGCCTGATCTGGGAACTCCTGCAGGCCTCTGCCTTCTGTGGGTTAGGCTT CCTGATAGTGCTAGCTCTCTTTCAGGCAGGGTTGGGTAGAATGATGATGAAGTACAG AGACCAGAGGGCTGGGAAGATATCTGAGAGGCTGGTCATTACTTCTGAAATGATAG AAAACATCCAGTCTGTTAAAGCTTACTGCTGGGAGGAGGCTATGGAAAAGATGATT GAGAACTTGAGGCAAACAGAGCTCAAGCTGACTAGGAAGGCAGCCTATGTCAGGTA TITTCAACAGCAGTGCTTTCTTCTTCTCAGGCTTTTTCGTGGTCTTCTTGAGTGITCTGC CCTATGCCCTCATCAAGGGGATAATTTTGAGAAAGATTTTCACCACTATTTICCTTTIG CATTGTCCTGAGGATGGCTGTCACCAGGCAATTCCCCTGGGCTGTGCAGACATGGTA TGACTCTCTGGGGGCCATCAACAAAATCCAAGATTTCCTGCAGAAGCAGGAGTACA AGACCCTGGAATACAACCTCACCACCACAGAAGTTIGTGATGGAGAATGTGACTGCA TTCTGGGAGGAAGGATTTGGGGAGCTGTTTGAGAAAGCAAAACAAAACAATAATAA CAGGAAAACCAGCAATGGAGATGACTCCCTGTTCTTTTCCAACTTCTCTTTGTTGGG CACCCCTGTCCTGAAAGATATAAACTTTAAAATTGAAAGAGGGCAGCTGTTGGCAGT TGCTGGCTCCACAGGAGCTGGAAAAACTTCACTACTGATGGTGATCATGGGGGAGTT AGAACCCTCTGAAGGGAAAATAAAACATTCTGGGAGGATTAGTTTCTGCAGCCAGT TCAGCTGGATCATGCCTGGGACCATTAAAGAAAATATTATATTTGGAGTGAGCTATG ATGAATATAGATATAGGAGTGTCATCAAAGCCTGTCAGTTGGAGGAAGACATCAGC AAATTTGCAGAGAAAGACAACATTGTTCTGGGTGAAGGTGGCATCACCCTGTCAGG AGGGCAAAGGGCCAGGATCAGCTTGGCCAGAGCAGTCTATAAAGATGCTGATCTGT ACCTCCTGGATAGCCCTTTTGGCTATCTGGATGTTTTGACAGAGAAGGAAATTTTTG AGTCCTGTGTCTGCAAGTTAATGGCAAATAAAACAAGGATACTTGTGACCTCAAAA ATGGAACACCTGAAGAAGGCTGACAAAATTCTGATCCTGCATGAGGGCAGCAGCTA CTTTTATGGAACATTTTCTGAACTGCAGAATTTGCAACCAGACTTTTCATCAAAGCTC ATGGGATGTGACAGTTTTGATCAGTTTTCTGCAGAAAGGAGAAACTCCATTTTGACT GAGACCCTGCACAGGTTCAGTCTGGAGGGGGATGCCCCAGTGAGTTGGACTGAGAC AAAGAAACAGAGCTTCAAGCAGACTGGAGAGTTTGGAGAAAAGAGGAAAAACTCA ATTCTCAATCCCATCAATAGCATCAGGAAGTTCAGCATAGTTCAGAAGACTCCTITG CAGATGAATGGGATTGAAGAGGACTCAGATGAGCCCCTGGAAAGGAGACTCTCCTT GGTGCCAGATTCAGAGCAGGGGGAAGCCATACTGCCAAGGATCTCTGTGATTTCTAC AGGGCCCACCCTCCAAGCAAGAAGGAGACAGTCAGTTTTAAACCTGATGACCCACT CTGTCAACCAGGGACAGAACATTCATAGAAAGACAACAGCATCTACAAGAAAAGTT TCACTGGCCCCTCAAGCCAATTTAACTGAACTAGATATCTACAGCAGGAGGCTCAGC CAAGAAACAGGCCTGGAGATCTCAGAAGAAATAAATGAGGAGGATTTGAAGGAAT GCTTCTTTGATGATATGGAGAGCATCCCAGCTGTCACAACCTGGAACACCTACCTGA GATACATCACAGTGCACAAATCCCTCATCTTTGTACTTATATGGTGCCTTGTCATCTT CTTAGCTGAGGTGGCTGCTTCCCTGGTGGTGCTGTGGCTGCTGGGAAACACACCCCT CCAGGATAAAGGGAACTCTACTCACAGCAGGAACAACAGTTATGCTGTGATCATCA CCAGTACCTCCTCCTACTATGTGTTCTACATTTATGTTGGAGTTGCAGACACATTGCT TGCCATGGGTTTTTTTAGAGGACTCCCCCTGGTGCATACTCTCATCACTGTTTCCAAA ATCCTTCACCACAAGATGCTGCACAGTGTACTACAGGCTCCCATGAGCACCCTCAAC ACTCTTAAAGCAGGAGGAATCTTGAACAGATTTAGCAAGGACATTGCAATTCTTGAT GACCTGCTTCCACTGACCATCTTTGACTTCATCCAGCTTCTGCTCATTGTAATTGGTG CCATTGCTGTGGTAGCAGTGCTCCAGCCATATATTTTTGTGGCCACTGTGCCTGTTAT TGTGGCCTTCATTATGTTGAGAGCCTACTTCCTGCAGACCTCTCAGCAGCTCAAGCA ACTTGAAAGTGAGGGCAGGAGCCCCATATTTACACACTTGGTCACTTCCCTCAAAGG CCTCTGGACACTCAGAGCTTTTGGAAGACAACCTTATTTTGAAACTCTCTTCCACAA GGCTCTGAATCTCCACACAGCCAACTGGTTTCTGTATCTTTCAACACTGCGCTGGTTC CAGATGAGGATTGAGATGATCTTTGTTATCTTCTTCATAGCTGTTACCTTCATCTCTA TTCTGACAACTGGTGAGGGGGAAGGGAGAGTAGGCATCATCCTCACACTAGCCATG AACATAATGTCTACCTTACAATGGGCCGTGAACAGCTCCATAGATGTGGACAGCCTC ATGAGAAGTGTGTCAAGAGTTTTCAAATTCATTGACATGCCCACAGAAGGCAAACC AACCAAGAGCACAAAACCCTACAAGAATGGCCAGCTGAGTAAGGTCATGATCATTG AAAATTCTCATGTGAAGAAGGATGATATTTGGCCCAGTGGGGGCCAGATGACAGTC AAGGACCTCACTGCCAAATACACAGAGGGTGGAAATGCTATCCTAGAGAACATCTC CTTCTCCATCTCCCCAGGCCAAAGAGTTGGCTTGCTGGGCAGGACTGGCAGTGGCAA GTCCACCTTGCTCTCAGCATTTCTCAGGCTTTTAAATACAGAGGGAGAGATTCAAAT TGATGGGGTGTCTTGGGATAGTATAACACTTCAACAGTGGAGGAAAGCCTTTGGTGT GATTCCTCAGAAAGTGTTTATCTTCTCTGGCACTTTCAGAAAAAATCTGGACCCCTAT GAACAGTGGAGTGACCAGGAAATCTGGAAGGTGGCAGATGAAGTGGGCCTAAGATC AGTCATAGAGCAGTTTCCTGGAAAGTTGGATTTTGTGCTTGTAGATGGAGGCTGTGT GCTGTCCCATGGCCATAAACAGCTAATGTGCCTGGCTAGGTCAGTGCTGAGCAAGGC CAAGATCCTGCTGTTAGATGAGCCTTCAGCCCATCTGGACCCTGTGACATACCAGAT TATCAGAAGAACTCTGAAGCAGGCCTTTGCTGACTGCACTGTCATCCTGTGTGAGCA CAGAATTGAGGCCATGCTGGAGTGCCAGCAGTTCCTTGTTATAGAAGAGAATAAGG TTAGGCAGTATGACAGCATTCAGAAACTGCTAAATGAAAGATCTCTCTTCAGGCAAG CTATTTCACCATCTGATAGAGTGAAACTTTTTCCCCACAGAAATTCCTCTAAATGTAA ATCTAAGCCCCAGATAGCTGCCTTGAAAGAGGAGACTGAAGAAGAAGTCCAGGACA CCAGACTGTGA (SEQ ID NO: 33) ATGCAGAGATCCCCGCTGGAGAAGGCATCTGTGGTGTCAAAACTGTTCTTTAGCTGG ACAAGGCCCATCCTTAGGAAAGGGTACAGACAGAGGTTGGAGCTGTCAGACATATA TCAGATCCCTTCAGTGGACTCTGCAGACAACCTCTCTGAAAAGCTGGAGAGGGAAT GGGACAGGGAACTGGCCAGCAAAAAAAACCCTAAACTGATTAATGCCCTGAGGAGG TGCTTCTTTTGGAGATTCATGTTCTATGGGATCTTCCTTTACCTGGGGGAGGTGACTA AAGCTGTTCAGCCTCTTCTTCTGGGGAGGATTATTGCCTCCTATGACCCAGACAACA AAGAAGAAAGAAGCATAGCCATTTACTTAGGCATAGGCCTCTGCTTGCTCTTCATAG TTAGAACCCTCCTACTCCACCCAGCCATCTTTGGTCTCCACCACATAGGTATGCAGA TGAGAATAGCAATGTTCTCCTTGATCTACAAGAAGACCCTCAAGCTGTCCAGCAGGG TGCTGGACAAGATCTCCATAGGCCAGTTAGTCAGTCTACTGTCCAATAACTTAAATA AGTTTGATGAGGGACTGGCACTGGCACATTTTGTGTGGATTGCCCCCCTCCAAGTGG CCCTTCTTATGGGCCTTATCTGGGAGCTGTTGCAGGCCTCTGCTTTCTGTGGCCTGGG TTTCCTCATAGTCCTAGCCTTATTCCAGGCTGGACTGGGCAGAATGATGATGAAGTA TAGGGACCAAAGAGCAGGGAAGATTTCTGAAAGGCTGGTTATAACTTCTGAGATGA TTGAGAACATTCAGTCAGTGAAAGCTTACTGCTGGGAAGAAGCTATGGAAAAAATG ATTGAAAATCTCAGACAGACTGAATTAAAGTTGACCAGGAAAGCTGCTTATGTCAG ATACTTCAACTCCTCAGCCTTCTTTTTTTCTGGCTTCTTTGTTGTATTCCTTTCAGTCC TCCCCTATGCCCTGATTAAGGGCATTATCTTGAGGAAAATTTTCACAACCATCTCCTT TTGTATTGTCCTCAGGATGGCTGTTACAAGGCAATTTCCTTGGGCTGTGCAAACTTG GTATGATAGCCTTGGAGCAATCAACAAGATCCAGGATTTCCTGCAAAAGCAGGAGT ACAAGACATTGGAATACAACCTTACCACCACTGAGGTGGTGATGGAAAATGTGACT GCCTTCTGGGAGGAGGGGTTTGGAGAGCTGTTTGAGAAAGCCAAACAGAACAACAA CAATAGAAAGACCTCTAATGGTGATGATTCCCTGTTCTTTTCTAACTTTAGTCTICTG GGGACCCCAGTTCTGAAAGATATTAACTTTAAAATTGAAAGGGGACAGTTGCTGGCT GTGGCTGGGTCCACTGGGGCTGGGAAGACAAGCCTGCTCATGGTGATCATGGGAGA GCTGGAACCCAGTGAAGGAAAGATCAAACACTCAGGCAGGATCTCCTTCTGCAGCC AGTTCTCATGGATTATGCCAGGCACTATTAAAGAAAATATCATCTTTGGTGTAAGCT ATGATGAGTACAGGTATAGATCTGTAATTAAAGCCTGCCAGCTGGAGGAAGACATC TCTAAGTTTGCTGAGAAGGATAACATTGTGTTGGGGGAAGGGGGCATCACCCTTTCT GGTGGGCAGAGGGCTAGGATCTCCCTTGCTAGGGCAGTATACAAGGATGCTGACTT GTACCTCTTGGATAGTCCTTTTGGCTACCTAGATGTGCTGACAGAGAAAGAAATATT TGAAAGCTGTGTGTGTAAGCTCATGGCTAACAAGACCAGGATCCTGGTCACCAGTA AAATGGAACACCTCAAAAAAGCAGACAAGATCCTTATTCTCCATGAGGGCTCCTCCT ACTTCTATGGGACCTTCAGTGAGCTGCAGAATCTGCAGCCAGACTTCTCCTCAAAAC TTATGGGCTGTGACTCCTTTGACCAATTCTCTGCAGAAAGAAGGAATAGCATACTGA CAGAAACACTGCATAGATTCTCCCTGGAAGGAGATGCCCCAGTGAGTTGGACAGAA ACCAAAAAGCAGAGCTTCAAGCAGACTGGTGAGTTTGGTGAAAAGAGGAAGAATTC TATCCTGAACCCCATCAATAGCATCAGGAAATTTAGCATAGTCCAAAAGACCCCCCT CCAGATGAATGGAATAGAGGAGGATAGTGATGAGCCTCTTGAGAGAAGGCTGTCCC TGGTTCCAGACAGTGAACAGGGTGAAGCCATTCTTCCGAGGATCAGTGTCATCTCCA CTGGGCCCACATTGCAGGCCAGAAGAAGACAGTCTGTTCTGAATTTGATGACACATT CTGTGAATCAAGGCCAGAATATCCATAGAAAAACCACTGCCAGCACCAGAAAAGTT TCTCTAGCCCCCCAGGCTAACCTGACTGAGTTAGACATCTACAGCAGAAGGCTGAGC CAAGAGACTGGCTTGGAAATATCTGAGGAGATCAATGAGGAGGACCTCAAGGAGTG CTTCTTTGATGACATGGAGTCAATCCCTGCAGTCACTACATGGAACACTTACCTAAG GTACATCACAGTTCATAAGAGCCTCATCTTTGTCCTCATATGGTGTCTGGTCATCTTT TTAGCAGAAGTGGCTGCCAGCCTAGTTGTGCTGTGGTTACTGGGCAATACACCTCTT CAGGACAAAGGCAATAGCACACACAGCAGAAACAACTCCTATGCAGTGATCATCAC CTCTACAAGCTCTTACTATGTATTCTATATATATGTGGGAGTGGCAGATACTCTCCTG GCCATGGGATTCTTCAGGGGATTACCTCTAGTTCACACATTGATCACAGTGTCAAAA ATTCTCCACCACAAGATGTTACACAGTGTCCTGCAAGCCCCAATGTCTACTCTGAAC ACACTTAAGGCAGGTGGAATTTTGAATAGGTTTAGCAAGGACATAGCTATCCTGGAT GATCTCCTCCCTCTGACCATCTITGACTTCATCCAGTTACTGCTCATTGTAATTGGAG CCATTGCAGTGGTAGCAGTCCTACAGCCTTACATTTTTGTGGCTACTGTTCCTGTTAT TGTGGCCTTCATTATGCTAAGAGCTTACTTCCTGCAAACAAGCCAACAGTTGAAACA GCTAGAAAGTGAGGGAAGGTCCCCCATCTTCACCCACCTGGTGACATCACTCAAGG GGCTATGGACTCTTAGGGCTTTTGGGAGACAGCCGTACTTTGAGACCTTATTCCATA AGGCCCTTAACCTCCATACAGCAAACTGGTTCTTATACCTGAGTACTCTGAGGTGGT TTCAAATGAGGATTGAAATGATTTTTGTGATCTTCTTCATTGCTGTGACCTTCATCTC AATCTTGACCACAGGAGAGGGGGAGGGCAGGGTGGGCATCATACTGACCTTGGCCA TGAACATTATGTCAACCCTGCAGTGGGCTGTCAATAGCTCCATTGATGTGGACAGTC TGATGAGGAGTGTCTCCAGGGTCTTCAAGTTTATTGACATGCCAACTGAGGGCAAAC CCACCAAAAGCACTAAGCCATATAAAAATGGCCAACTGTCCAAAGTGATGATCATT GAAAATTCACATGTAAAGAAGGATGATATCTGGCCCTCTGGAGGACAGATGACAGT GAAAGACCTGACTGCCAAGTACACAGAGGGTGGTAATGCCATTCTTGAGAACATTA GTTTCAGTATTTCCCCGGGGCAAAGGGTGGGCCTCCTTGGCAGAACAGGCTCTGGCA AGAGTACCCTGCTGTCAGCCTTTTTAAGACTGTTGAACACTGAGGGAGAAATTCAGA TTGATGGTGTCTCCTGGGATAGCATCACCCTCCAGCAGTGGAGAAAAGCTTTITGGAG TGATCCCGCAAAAGGTTTTCATCTTTTCAGGCACCTTCCGGAAGAACCTGGACCCCT ATGAGCAGTGGTCTGACCAGGAAATATGGAAGGTAGCTGATGAAGTTGGGCTTAGG TCAGTCATAGAGCAGTTCCCAGGCAAACTGGACTTTGTCCTGGTGGATGGTGGATGT GTACTGAGTCATGGGCACAAACAGCTGATGTGCCTAGCCAGGTCTGTGCTCAGCAA GGCAAAGATATTGCTGCTTGATGAACCCAGTGCCCATCTGGACCCAGTCACATATCA GATCATCAGAAGAACATTGAAGCAGGCCTTTGCTGATTGCACAGTTATCCTCTGTGA GCACAGGATTGAGGCCATGCTGGAGTGCCAGCAGTTTCTGGTGATTGAGGAGAATA AAGTAAGGCAGTATGACTCCATCCAGAAGCTGCTCAATGAAAGAAGCCTCTTTAGA CAAGCTATCTCCCCCTCAGACAGGGTCAAATTGTTCCCTCACAGAAACAGCAGCAA GTGCAAGAGCAAGCCCCAAATTGCAGCCTTGAAAGAGGAGACAGAGGAAGAGGTG CAGGACACCAGACTCTGA (SEQ ID NO: 34) ATGCAGAGAAGCCCCCTGGAGAAGGCCAGCGTGGTGAGCAAGCTGTTCTTCAGCTG GACCAGACCCATCCTGAGAAAGGGCTACAGACAGAGACTGGAGCTGAGCGACATCT ACCAGATCCCCAGCGTGGACAGCGCCGACAACCTGAGCGAGAAGCTGGAGAGAGA GTGGGACAGAGAGCTGGCCAGCAAGAAGAACCCCAAGCTGATCAACGCCCTGAGA AGATGCTTCTTCTGGAGATTCATGTTCTACGGCATCTTCCTGTACCTGGGCGAGGTG ACCAAGGCCGTGCAGCCCCTGCTGCTGGGCAGAATCATCGCCAGCTACGACCCCGA CAACAAGGAGGAGAGAAGCATCGCCATCTACCTGGGCATCGGCCTGTGCCTGCTGT TCATCGTGAGAACCCTGCTGCTGCACCCCGCCATCTTCGGCCTGCACCACATCGGCA TGCAGATGAGAATCGCCATGTTCAGCCTGATCTACAAGAAGACCCTGAAGCTGAGC AGCAGAGTGCTGGACAAGATCAGCATCGGCCAGCTGGTGAGCCTGCTGAGCAACAA CCTGAACAAGTTCGACGAGGGCCTGGCCCTGGCCCACTTCGTGTGGATCGCCCCCCT GCAGGTGGCCCTGCTGATGGGCCTGATCTGGGAGCTGCTGCAGGCCAGCGCCTICTG CGGCCTGGGCTTCCTGATCGTGCTGGCCCTGTTCCAGGCCGGCCTGGGCAGAATGAT GATGAAGTACAGAGACCAGAGAGCCGGCAAGATCAGCGAGAGACTGGTGATCACC AGCGAGATGATCGAGAACATCCAGAGCGTGAAGGCCTACTGCTGGGAGGAGGCCAT GGAGAAGATGATCGAGAACCTGAGACAGACCGAGCTGAAGCTGACCAGAAAGGCC GCCTACGTGAGATACTTCAACAGCAGCGCCTTCTTCTTCAGCGGCTTCTTCGTGGTGT TCCTGAGCGTGCTGCCCTACGCCCTGATCAAGGGCATCATCCTGAGAAAGATCTTCA CCACCATCAGCTTCTGCATCGTGCTGAGAATGGCCGTGACCAGACAGTTCCCCTGGG CCGTGCAGACCTGGTACGACAGCCTGGGCGCCATCAACAAGATCCAGGACTTCCTG CAGAAGCAGGAGTACAAGACCCTGGAGTACAACCTGACCACCACCGAGGTGGTGAT GGAGAACGTGACCGCCTTCTGGGAGGAGGGCTTCGGCGAGCTGTTCGAGAAGGCCA AGCAGAACAACAACAACAGAAAGACCAGCAACGGCGACGACAGCCTGTTCTTCAGC AACTTCAGCCTGCTGGGCACCCCCGTGCTGAAGGACATCAACTTCAAGATCGAGAG AGGCCAGCTGCTGGCCGTGGCCGGCAGCACCGGCGCCGGCAAGACCAGCCTGCTGA TGGTGATCATGGGCGAGCTGGAGCCCAGCGAGGGCAAGATCAAGCACAGCGGCAG AATCAGCTTCTGCAGCCAGTTCAGCTGGATCATGCCCGGCACCATCAAGGAGAACA TCATCTTCGGCGTGAGCTACGACGAGTACAGATACAGAAGCGTGATCAAGGCCTGC CAGCTGGAGGAGGACATCAGCAAGTTCGCCGAGAAGGACAACATCGTGCTGGGCGA GGGCGGCATCACCCTGAGCGGCGGCCAGAGAGCCAGAATCAGCCTGGCCAGAGCCG TGTACAAGGACGCCGACCTGTACCTGCTGGACAGCCCCTTCGGCTACCTGGACGTGC TGACCGAGAAGGAGATCTTCGAGAGCTGCGTGTGCAAGCTGATGGCCAACAAGACC AGAATCCTGGTGACCAGCAAGATGGAGCACCTGAAGAAGGCCGACAAGATCCTGAT CCTGCACGAGGGCAGCAGCTACTTCTACGGCACCTTCAGCGAGCTGCAGAACCTGC AGCCCGACTTCAGCAGCAAGCTGATGGGCTGCGACAGCTTCGACCAGTTCAGCGCC GAGAGAAGAAACAGCATCCTGACCGAGACCCTGCACAGATTCAGCCTGGAGGGCGA CGCCCCCGTGAGCTGGACCGAGACCAAGAAGCAGAGCTTCAAGCAGACCGGCGAGT TCGGCGAGAAGAGAAAGAACAGCATCCTGAACCCCATCAACAGCATCAGAAAGTTC AGCATCGTGCAGAAGACCCCCCTGCAGATGAACGGCATCGAGGAGGACAGCGACG AGCCCCTGGAGAGAAGACTGAGCCTGGTGCCCGACAGCGAGCAGGGCGAGGCCATC CTGCCCAGAATCAGCGTGATCAGCACCGGCCCCACCCTGCAGGCCAGAAGAAGACA GAGCGTGCTGAACCTGATGACCCACAGCGTGAACCAGGGCCAGAACATCCACAGAA AGACCACCGCCAGCACCAGAAAGGTGAGCCTGGCCCCCCAGGCCAACCTGACCGAG CTGGACATCTACAGCAGAAGACTGAGCCAGGAGACCGGCCTGGAGATCAGCGAGG AGATCAACGAGGAGGACCTGAAGGAGTGCTTCTTCGACGACATGGAGAGCATCCCC GCCGTGACCACCTGGAACACCTACCTGAGATACATCACCGTGCACAAGAGCCTGAT CTTCGTGCTGATCTGGTGCCTGGTGATCTTCCTGGCCGAGGTGGCCGCCAGCCTGGT GGTGCTGTGGCTGCTGGGCAACACCCCCCTGCAGGACAAGGGCAACAGCACCCACA GCAGAAACAACAGCTACGCCGTGATCATCACCAGCACCAGCAGCTACTACGTGTTC TACATCTACGTGGGCGTGGCCGACACCCTGCTGGCCATGGGCTTCTTCAGAGGCCTG CCCCTGGTGCACACCCTGATCACCGTGAGCAAGATCCTGCACCACAAGATGCTGCAC AGCGTGCTGCAGGCCCCCATGAGCACCCTGAACACCCTGAAGGCCGGCGGCATCCT GAACAGATTCAGCAAGGACATCGCCATCCTGGACGACCTGCTGCCCCTGACCATCTT CGACTTCATCCAGCTGCTGCTGATCGTGATCGGCGCCATCGCCGTGGTGGCCGTGCT GCAGCCCTACATCTTCGTGGCCACCGTGCCCGTGATCGTGGCCTTCATCATGCTGAG AGCCTACTTCCTGCAGACCAGCCAGCAGCTGAAGCAGCTGGAGAGCGAGGGCAGAA GCCCCATCTTCACCCACCTGGTGACCAGCCTGAAGGGCCTGTGGACCCTGAGAGCCT TCGGCAGACAGCCCTACTTCGAGACCCTGTTCCACAAGGCCCTGAACCTGCACACCG CCAACTGGTTCCTGTACCTGAGCACCCTGAGATGGTTCCAGATGAGAATCGAGATGA TCTTCGTGATCTTCTTCATCGCCGTGACCTTCATCAGCATCCTGACCACCGGCGAGG GCGAGGGCAGAGTGGGCATCATCCTGACCCTGGCCATGAACATCATGAGCACCCTG CAGTGGGCCGTGAACAGCAGCATCGACGTGGACAGCCTGATGAGAAGCGTGAGCAG AGTGTTCAAGTTCATCGACATGCCCACCGAGGGCAAGCCCACCAAGAGCACCAAGC CCTACAAGAACGGCCAGCTGAGCAAGGTGATGATCATCGAGAACAGCCACGTGAAG AAGGACGACATCTGGCCCAGCGGCGGCCAGATGACCGTGAAGGACCTGACCGCCAA GTACACCGAGGGCGGCAACGCCATCCTGGAGAACATCAGCTTCAGCATCAGCCCCG GCCAGAGAGTGGGCCTGCTGGGCAGAACCGGCAGCGGCAAGAGCACCCTGCTGAGC GCCTTCCTGAGACTGCTGAACACCGAGGGCGAGATCCAGATCGACGGCGTGAGCTG GGACAGCATCACCCTGCAGCAGTGGAGAAAGGCCTTCGGCGTGATCCCCCAGAAGG TGTTCATCTTCAGCGGCACCTTCAGAAAGAACCTGGACCCCTACGAGCAGTGGAGC GACCAGGAGATCTGGAAGGTGGCCGACGAGGTGGGCCTGAGAAGCGTGATCGAGC AGTTCCCCGGCAAGCTGGACTTCGTGCTGGTGGACGGCGGCTGCGTGCTGAGCCAC GGCCACAAGCAGCTGATGTGCCTGGCCAGAAGCGTGCTGAGCAAGGCCAAGATCCT GCTGCTGGACGAGCCCAGCGCCCACCTGGACCCCGTGACCTACCAGATCATCAGAA GAACCCTGAAGCAGGCCTTCGCCGACTGCACCGTGATCCTGTGCGAGCACAGAATC GAGGCCATGCTGGAGTGCCAGCAGTTCCTGGTGATCGAGGAGAACAAGGTGAGACA GTACGACAGCATCCAGAAGCTGCTGAACGAGAGAAGCCTGTTCAGACAGGCCATCA GCCCCAGCGACAGAGTGAAGCTGTTCCCCCACAGAAACAGCAGCAAGTGCAAGAGC AAGCCCCAGATCGCCGCCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCA GACTGTGA (SEQ ID NO: 35) ATGCAGCGCAGCCCCCTGGAGAAGGCCAGCGTGGTGAGCAAGCTGTTCTTCAGCTG GACCCGCCCCATCCTGCGCAAGGGCTACCGCCAGCGCCTGGAGCTGAGCGACATCT ACCAGATCCCCAGCGTGGACAGCGCCGACAACCTGAGCGAGAAGCTGGAGCGCGA GTGGGACCGCGAGCTGGCCAGCAAGAAGAACCCCAAGCTGATCAACGCCCTGCGCC GCTGCTTCTTCTGGCGCTTCATGTTCTACGGCATCTTCCTGTACCTGGGCGAGGTGAC CAAGGCCGTGCAGCCCCTGCTGCTGGGCCGCATCATCGCCAGCTACGACCCCGACA ACAAGGAGGAGCGCAGCATCGCCATCTACCTGGGCATCGGCCTGTGCCTGCTGTTCA TCGTGCGCACCCTGCTGCTGCACCCCGCCATCTTCGGCCTGCACCACATCGGCATGC AGATGCGCATCGCCATGTTCAGCCTGATCTACAAGAAGACCCTGAAGCTGAGCAGC CGCGTGCTGGACAAGATCAGCATCGGCCAGCTGGTGAGCCTGCTGAGCAACAACCT GAACAAGTTCGACGAGGGCCTGGCCCTGGCCCACTTCGTGTGGATCGCCCCCCTGCA GGTGGCCCTGCTGATGGGCCTGATCTGGGAGCTGCTGCAGGCCAGCGCCTTCTGCGG CCTGGGCTTCCTGATCGTGCTGGCCCTGTTCCAGGCCGGCCTGGGCCGCATGATGAT GAAGTACCGCGACCAGCGCGCCGGCAAGATCAGCGAGCGCCTGGTGATCACCAGCG AGATGATCGAGAACATCCAGAGCGTGAAGGCCTACTGCTGGGAGGAGGCCATGGAG AAGATGATCGAGAACCTGCGCCAGACCGAGCTGAAGCTGACCCGCAAGGCCGCCTA CGTGCGCTACTTCAACAGCAGCGCCTTCTTCTTCAGCGGCTTCTTCGTGGTGTICCTG AGCGTGCTGCCCTACGCCCTGATCAAGGGCATCATCCTGCGCAAGATCTTCACCACC ATCAGCTTCTGCATCGTGCTGCGCATGGCCGTGACCCGCCAGTTCCCCTGGGCCGTG CAGACCTGGTACGACAGCCTGGGCGCCATCAACAAGATCCAGGACTTCCTGCAGAA GCAGGAGTACAAGACCCTGGAGTACAACCTGACCACCACCGAGGTGGTGATGGAGA ACGTGACCGCCTTCTGGGAGGAGGGCTTCGGCGAGCTGTTCGAGAAGGCCAAGCAG AACAACAACAACCGCAAGACCAGCAACGGCGACGACAGCCTGTTCTTCAGCAACTT CAGCCTGCTGGGCACCCCCGTGCTGAAGGACATCAACTTCAAGATCGAGCGCGGCC AGCTGCTGGCCGTGGCCGGCAGCACCGGCGCCGGCAAGACCAGCCTGCTGATGGTG ATCATGGGCGAGCTGGAGCCCAGCGAGGGCAAGATCAAGCACAGCGGCCGCATCA GCTTCTGCAGCCAGTTCAGCTGGATCATGCCCGGCACCATCAAGGAGAACATCATCT TCGGCGTGAGCTACGACGAGTACCGCTACCGCAGCGTGATCAAGGCCTGCCAGCTG GAGGAGGACATCAGCAAGTTCGCCGAGAAGGACAACATCGTGCTGGGCGAGGGCG GCATCACCCTGAGCGGCGGCCAGCGCGCCCGCATCAGCCTGGCCCGCGCCGTGTAC AAGGACGCCGACCTGTACCTGCTGGACAGCCCCTTCGGCTACCTGGACGTGCTGACC GAGAAGGAGATCTTCGAGAGCTGCGTGTGCAAGCTGATGGCCAACAAGACCCGCAT CCTGGTGACCAGCAAGATGGAGCACCTGAAGAAGGCCGACAAGATCCTGATCCTGC ACGAGGGCAGCAGCTACTTCTACGGCACCTTCAGCGAGCTGCAGAACCTGCAGCCC GACTTCAGCAGCAAGCTGATGGGCTGCGACAGCTTCGACCAGTTCAGCGCCGAGCG CCGCAACAGCATCCTGACCGAGACCCTGCACCGCTTCAGCCTGGAGGGCGACGCCC CCGTGAGCTGGACCGAGACCAAGAAGCAGAGCTTCAAGCAGACCGGCGAGTTCGGC GAGAAGCGCAAGAACAGCATCCTGAACCCCATCAACAGCATCCGCAAGTTCAGCAT CGTGCAGAAGACCCCCCTGCAGATGAACGGCATCGAGGAGGACAGCGACGAGCCCC TGGAGCGCCGCCTGAGCCTGGTGCCCGACAGCGAGCAGGGCGAGGCCATCCTGCCC CGCATCAGCGTGATCAGCACCGGCCCCACCCTGCAGGCCCGCCGCCGCCAGAGCGT GCTGAACCTGATGACCCACAGCGTGAACCAGGGCCAGAACATCCACCGCAAGACCA CCGCCAGCACCCGCAAGGTGAGCCTGGCCCCCCAGGCCAACCTGACCGAGCTGGAC ATCTACAGCCGCCGCCTGAGCCAGGAGACCGGCCTGGAGATCAGCGAGGAGATCAA CGAGGAGGACCTGAAGGAGTGCTTCTTCGACGACATGGAGAGCATCCCCGCCGTGA CCACCTGGAACACCTACCTGCGCTACATCACCGTGCACAAGAGCCTGATCTTCGTGC TGATCTGGTGCCTGGTGATCTTCCTGGCCGAGGTGGCCGCCAGCCTGGTGGTGCTGT GGCTGCTGGGCAACACCCCCCTGCAGGACAAGGGCAACAGCACCCACAGCCGCAAC AACAGCTACGCCGTGATCATCACCAGCACCAGCAGCTACTACGTGTTCTACATCTAC GTGGGCGTGGCCGACACCCTGCTGGCCATGGGCTTCTICCGCGGCCTGCCCCTGGTG CACACCCTGATCACCGTGAGCAAGATCCTGCACCACAAGATGCTGCACAGCGTGCT GCAGGCCCCCATGAGCACCCTGAACACCCTGAAGGCCGGCGGCATCCTGAACCGCT TCAGCAAGGACATCGCCATCCTGGACGACCTGCTGCCCCTGACCATCTTCGACTTCA TCCAGCTGCTGCTGATCGTGATCGGCGCCATCGCCGTGGTGGCCGTGCTGCAGCCCT ACATCTTCGTGGCCACCGTGCCCGTGATCGTGGCCTTCATCATGCTGCGCGCCTACTT CCTGCAGACCAGCCAGCAGCTGAAGCAGCTGGAGAGCGAGGGCCGCAGCCCCATCT TCACCCACCTGGTGACCAGCCTGAAGGGCCTGTGGACCCTGCGCGCCTTCGGCCGCC AGCCCTACTTCGAGACCCTGTTCCACAAGGCCCTGAACCTGCACACCGCCAACTGGT TCCTGTACCTGAGCACCCTGCGCTGGTTCCAGATGCGCATCGAGATGATCTTCGTGA TCTTCTTCATCGCCGTGACCTTCATCAGCATCCTGACCACCGGCGAGGGCGAGGGCC GCGTGGGCATCATCCTGACCCTGGCCATGAACATCATGAGCACCCTGCAGTGGGCC GTGAACAGCAGCATCGACGTGGACAGCCTGATGCGCAGCGTGAGCCGCGTGTTCAA GTTCATCGACATGCCCACCGAGGGCAAGCCCACCAAGAGCACCAAGCCCTACAAGA ACGGCCAGCTGAGCAAGGTGATGATCATCGAGAACAGCCACGTGAAGAAGGACGA CATCTGGCCCAGCGGCGGCCAGATGACCGTGAAGGACCTGACCGCCAAGTACACCG AGGGCGGCAACGCCATCCTGGAGAACATCAGCTTCAGCATCAGCCCCGGCCAGCGC GTGGGCCTGCTGGGCCGCACCGGCAGCGGCAAGAGCACCCTGCTGAGCGCCTTCCT GCGCCTGCTGAACACCGAGGGCGAGATCCAGATCGACGGCGTGAGCTGGGACAGCA TCACCCTGCAGCAGTGGCGCAAGGCCTTCGGCGTGATCCCCCAGAAGGTGTTCATCT TCAGCGGCACCTTCCGCAAGAACCTGGACCCCTACGAGCAGTGGAGCGACCAGGAG ATCTGGAAGGTGGCCGACGAGGTGGGCCTGCGCAGCGTGATCGAGCAGTTCCCCGG CAAGCTGGACTTCGTGCTGGTGGACGGCGGCTGCGTGCTGAGCCACGGCCACAAGC AGCTGATGTGCCTGGCCCGCAGCGTGCTGAGCAAGGCCAAGATCCTGCTGCTGGAC GAGCCCAGCGCCCACCTGGACCCCGTGACCTACCAGATCATCCGCCGCACCCTGAA GCAGGCCTTCGCCGACTGCACCGTGATCCTGTGCGAGCACCGCATCGAGGCCATGCT GGAGTGCCAGCAGTTCCTGGTGATCGAGGAGAACAAGGTGCGCCAGTACGACAGCA TCCAGAAGCTGCTGAACGAGCGCAGCCTGTTCCGCCAGGCCATCAGCCCCAGCGAC CGCGTGAAGCTGTITCCCCCACCGCAACAGCAGCAAGTGCAAGAGCAAGCCCCAGAT CGCCGCCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCCGCCTGTAA (SEQ ID NO: 36) ATGCAGAGAAGCCCCCTGGAGAAGGCCAGCGTGGTGAGCAAGCTGTTCTTCAGCTG GACCAGACCCATCCTGAGAAAGGGCTACAGACAGAGACTGGAGCTGAGCGACATCT ACCAGATCCCCAGCGTGGACAGCGCCGACAACCTGAGCGAGAAGCTGGAGAGAGA GTGGGACAGAGAGCTGGCCAGCAAGAAGAACCCCAAGCTGATCAACGCCCTGAGA AGATGCTITCTTCTIGGAGATTCATGTTCTACGGCATCTTCCTGTACCTGGGCGAGGTG ACCAAGGCCGTGCAGCCCCTGCTGCTGGGCAGAATCATCGCCAGCTACGACCCCGA CAACAAGGAGGAGAGAAGCATCGCCATCTACCTGGGCATCGGCCTGTGCCTGCTGT TCATCGTGAGAACCCTGCTGCTGCACCCCGCCATCTTCGGCCTGCACCACATCGGCA TGCAGATGAGAATCGCCATGTTCAGCCTGATCTACAAGAAGACCCTGAAGCTGAGC AGCAGAGTGCTGGACAAGATCAGCATCGGCCAGCTGGTGAGCCTGCTGAGCAACAA CCTGAACAAGTTCGACGAGGGCCTGGCCCTGGCCCACTTCGTGTGGATCGCCCCCCT GCAGGTGGCCCTGCTGATGGGCCTGATCTGGGAGCTGCTGCAGGCCAGCGCCTTICTG CGGCCTGGGCTTCCTGATCGTGCTGGCCCTGTTCCAGGCCGGCCTGGGCAGAATGAT GATGAAGTACAGGGACCAGAGAGCCGGCAAGATCAGCGAGAGACTGGTGATCACC AGCGAGATGATCGAGAACATCCAGAGCGTGAAGGCCTACTGCTGGGAGGAGGCCAT GGAGAAGATGATCGAGAACCTGAGACAGACCGAGCTGAAGCTGACCAGAAAGGCC GCCTACGTGAGATACTTCAACAGCAGCGCCTTCTTCTTCAGCGGCTTCTTCGTGGTGT TCCTGAGCGTGCTGCCCTACGCCCTGATCAAGGGCATCATCCTGAGAAAGATCTTCA CCACCATCAGCTTCTGCATCGTGCTGAGAATGGCCGTGACCAGACAGTTCCCCTGGG CCGTGCAGACCTGGTACGACAGCCTGGGCGCCATCAACAAGATCCAGGACTTCCTG CAGAAGCAGGAGTACAAGACCCTGGAGTACAACCTGACCACCACCGAGGTGGTGAT GGAGAACGTGACCGCCTTCTGGGAGGAGGGCTTCGGCGAGCTGTTCGAGAAGGCCA AGCAGAACAACAACAACAGAAAGACCAGCAACGGCGACGACAGCCTGTTCTTCAGC AACTTCAGCCTGCTGGGCACCCCCGTGCTGAAGGACATCAACTTCAAGATCGAGAG AGGCCAGCTGCTGGCCGTGGCCGGCAGCACCGGCGCCGGCAAGACCAGCCTGCTGA TGGTGATCATGGGCGAGCTGGAGCCCAGCGAGGGCAAGATCAAGCACAGCGGCAG AATCAGCTTCTGCAGCCAGTTCAGCTGGATCATGCCCGGCACCATCAAGGAGAACA TCATCTTCGGCGTGAGCTACGACGAGTACAGATACAGAAGCGTGATCAAGGCCTGC CAGCTGGAGGAGGACATCAGCAAGTTCGCCGAGAAGGACAACATCGTGCTGGGCGA GGGCGGCATCACCCTGAGCGGCGGCCAGAGAGCCAGAATCAGCCTGGCCAGAGCCG TGTACAAGGACGCCGACCTGTACCTGCTGGACAGCCCCTTCGGCTACCTGGACGTGC TGACCGAGAAGGAGATCTTCGAGAGCTGCGTGTGCAAGCTGATGGCCAACAAGACC AGAATCCTGGTGACCAGCAAGATGGAGCACCTGAAGAAGGCCGACAAGATCCTGAT CCTGCACGAGGGCAGCAGCTACTTCTACGGCACCTTCAGCGAGCTGCAGAACCTGC AGCCCGACTTCAGCAGCAAGCTGATGGGCTGCGACAGCTTCGACCAGTTCAGCGCC GAGAGAAGAAACAGCATCCTGACCGAGACCCTGCACAGATTCAGCCTGGAGGGCGA CGCCCCCGTGAGCTGGACCGAGACCAAGAAGCAGAGCTTCAAGCAGACCGGCGAGT TCGGCGAGAAGAGAAAGAACAGCATCCTGAACCCCATCAACAGCATCAGAAAGTTC AGCATCGTGCAGAAGACCCCCCTGCAGATGAACGGCATCGAGGAGGACAGCGACG AGCCCCTGGAGAGAAGACTGAGCCTGGTGCCCGACAGCGAGCAGGGCGAGGCCATC CTGCCCAGAATCAGCGTGATCAGCACCGGCCCCACCCTGCAGGCCAGAAGAAGACA GAGCGTGCTGAACCTGATGACCCACAGCGTGAACCAGGGCCAGAACATCCACAGAA AGACCACCGCCAGCACCAGAAAGGTGAGCCTGGCCCCCCAGGCCAACCTGACCGAG CTGGACATCTACAGCAGAAGACTGAGCCAGGAGACCGGCCTGGAGATCAGCGAGG AGATCAACGAGGAGGACCTGAAGGAGTGCTTCTTCGACGACATGGAGAGCATCCCC GCCGTGACCACCTGGAACACCTACCTGAGATACATCACCGTGCACAAGAGCCTGAT CTTCGTGCTGATCTGGTGCCTGGTGATCTTCCTGGCCGAGGTGGCCGCCAGCCTGGT GGTGCTGTGGCTGCTGGGCAACACCCCCCTGCAGGACAAGGGCAACAGCACCCACA GCAGAAACAACAGCTACGCCGTGATCATCACCAGCACCAGCAGCTACTACGTGTTC TACATCTACGTGGGCGTGGCCGACACCCTGCTGGCCATGGGCTTCTTCAGAGGCCTG CCCCTGGTGCACACCCTGATCACCGTGAGCAAGATCCTGCACCACAAGATGCTGCAC AGCGTGCTGCAGGCCCCCATGAGCACCCTGAACACCCTGAAGGCCGGCGGCATCCT GAACAGATTCAGCAAGGACATCGCCATCCTGGACGACCTGCTGCCCCTGACCATCTT CGACTTCATCCAGCTGCTGCTGATCGTGATCGGCGCCATCGCCGTGGTGGCCGTGCT GCAGCCCTACATCTTCGTGGCCACCGTGCCCGTGATCGTGGCCTTCATCATGCTGAG AGCCTACTTCCTGCAGACCAGCCAGCAGCTGAAGCAGCTGGAGAGCGAGGGCAGGA GCCCCATCTTCACCCACCTGGTGACCAGCCTGAAGGGCCTGTGGACCCTGAGAGCCT TCGGCAGACAGCCCTACTTCGAGACCCTGTTCCACAAGGCCCTGAACCTGCACACCG CCAACTGGTTCCTGTACCTGAGCACCCTGAGATGGTTCCAGATGAGAATCGAGATGA TCTTCGTGATCTTCTTCATCGCCGTGACCTTCATCAGCATCCTGACCACCGGCGAGG GCGAGGGCAGAGTGGGCATCATCCTGACCCTGGCCATGAACATCATGAGCACCCTG CAGTGGGCCGTGAACAGCAGCATCGACGTGGACAGCCTGATGAGAAGCGTGAGCAG AGTGTTCAAGTTCATCGACATGCCCACCGAGGGCAAGCCCACCAAGAGCACCAAGC CCTACAAGAACGGCCAGCTGAGCAAGGTGATGATCATCGAGAACAGCCACGTGAAG AAGGACGACATCTGGCCCAGCGGCGGCCAGATGACCGTGAAGGACCTGACCGCCAA GTACACCGAGGGCGGCAACGCCATCCTGGAGAACATCAGCTTCAGCATCAGCCCCG GCCAGAGAGTGGGCCTGCTGGGCAGAACCGGCAGCGGCAAGAGCACCCTGCTGAGC GCCTTCCTGAGACTGCTGAACACCGAGGGCGAGATCCAGATCGACGGCGTGAGCTG GGACAGCATCACCCTGCAGCAGTGGAGAAAGGCCTTCGGCGTGATCCCCCAGAAGG TGTTCATCTTCAGCGGCACCTTCAGAAAGAACCTGGACCCCTACGAGCAGTGGAGC GACCAGGAGATCTGGAAGGTGGCCGACGAGGTGGGCCTGAGAAGCGTGATCGAGC AGTTCCCCGGCAAGCTGGACTTCGTGCTGGTGGACGGCGGCTGCGTGCTGAGCCAC GGCCACAAGCAGCTGATGTGCCTGGCCAGAAGCGTGCTGAGCAAGGCCAAGATCCT GCTGCTGGACGAGCCCAGCGCCCACCTGGACCCCGTGACCTACCAGATCATCAGAA GAACCCTGAAGCAGGCCTTCGCCGACTGCACCGTGATCCTGTGCGAGCACAGAATC GAGGCCATGCTGGAGTGCCAGCAGTTCCTGGTGATCGAGGAGAACAAGGTGAGACA GTACGACAGCATCCAGAAGCTGCTGAACGAGAGAAGCCTGTTCAGACAGGCCATCA GCCCCAGCGACAGAGTGAAGCTGTTCCCCCACAGAAACAGCAGCAAGTGCAAGAGC AAGCCCCAGATCGCCGCCCTGAAGGAGGAGACCGAGGAGGAGGTGCAGGACACCA GACTGTGA (SEQ ID NO: 37) ATGCAGAGGTCACCTCTGGAAAAGGCTAGCGTGGTCAGCAAGCTATTTTTTTCCTGG ACCCGCCCGATACTCAGGAAGGGCTACCGACAGCGGCTGGAGCTGAGTGACATTTA TCAGATTCCCTCCGTCGATTCCGCTGACAACCTGTCTGAGAAACTGGAGCGGGAATG GGATAGGGAACTGGCGTCCAAAAAAAACCCCAAACTCATCAATGCACTCCGCAGAT GCTTCTTCTGGCGGTTTATGTTTTATGGCATATTCCTGTATCTGGGGGAGGTGACGAA AGCCGTGCAGCCGCTGCTGCTTGGTCGCATTATCGCGTCATACGATCCAGATAACAA GGAGGAAAGAAGTATCGCTATCTATCTCGGGATAGGGCTGTGCCTGCTCTTCATTGT GCGGACTCTTCTCTTIGCACCCCGCCATTTTCGGTCTGCATCATATAGGTATGCAGATG AGAATTGCGATGTTCTCATTGATTTACAAAAAAACGCTTAAGCTAAGTTCAAGGGTG CTAGATAAGATATCGATCGGCCAGCTGGTGTCTCTGCTTAGCAACAACCTCAATAAA TITCGACGAAGGCCTTGCACTGGCCCACTTCGTGTGGATCGCCCCTCTGCAGGTGGCT CTGCTGATGGGGTTAATATGGGAGCTGTTGCAGGCCTCCGCTTTTTGIGGCCTGGGG TITCTCATCGTGTTGGCCTTGTTTCAGGCAGGGCTGGGACGTATGATGATGAAATAT AGGGATCAGAGGGCTGGCAAAATCTCTGAGCGCCTGGTTATTACGAGTGAAATGAT TGAGAACATCCAGTCAGTGAAGGCCTATTGCTGGGAGGAGGCCATGGAAAAAATGA TTGAGAACCTACGCCAGACTGAGCTGAAGTTAACCAGAAAAGCCGCCTATGTGCGC TACTTTAACAGTAGCGCATTTTTCTTCTCCGGTTITTTCGTGGTGTTTCTTAGTGTGIT GCCGTATGCCTTAATCAAGGGAATAATACTCCGGAAGATTTTCACTACCATCAGCTT CTGTATCGTGTTGCGGATGGCCGTCACCCGGCAGTTTCCCTGGGCAGTACAGACTTG GTACGATTCTCTCGGAGCAATTAACAAAATCCAAGACTTTCTACAAAAGCAGGAGT ACAAGACCCTGGAGTACAATCTGACCACCACAGAAGTCGTAATGGAGAATGTAACT GCCTTCTGGGAAGAGGGCTTTGGCGAACTCTTTGAAAAGGCCAAGCAGAACAATAA CAACCGGAAGACCTCCAACGGGGACGACAGCTTATTTTTCAGCAATTTTTCTTTGCT CGGGACCCCTGTACTGAAAGATATTAACTTTAAGATCGAGCGCGGACAACTCCTGGC TGTCGCCGGCAGCACTGGAGCTGGAAAAACATCACTGCTTATGGTGATAATGGGAG AACTCGAACCAAGCGAGGGAAAAATAAAGCACTCTGGACGGATTAGTTTTTGCTCC CAGTTCTCGTGGATAATGCCTGGCACCATTAAGGAGAATATCATCTTTGGAGTGAGT TACGACGAATACCGGTACCGGTCCGTTATCAAGGCTTGTCAACTCGAGGAGGACATT TCTAAATTCGCCGAAAAAGATAATATAGTGCTGGGCGAAGGAGGCATTACACTGAG CGGGGGTCAGAGAGCTCGAATTAGCCTCGCCCGAGCAGTCTATAAAGACGCCGATC TTTACCTGCTGGATTCCCCTTTTGGGTATTTGGATGTTCTGACAGAGAAGGAAATCTT TGAATCATGTGTCTGTAAACTGATGGCCAATAAGACTAGGATTCTAGTGACTTCGAA AATGGAGCACCTGAAAAAAGCGGACAAAATTCTGATACTCCATGAAGGGTCTTCCT ACTTCTACGGCACCTTCTCAGAGTTGCAGAACTTACAACCTGATTTTTCATCTAAGCT TATGGGGTGCGACTCGTTTGACCAGTTCTCCGCTGAAAGACGAAACAGCATCTTAAC GGAAACTCTTCACAGGTTCTCATTAGAGGGAGATGCGCCGGTGTCCTGGACAGAGA CAAAAAAACAGTCTTTCAAACAGACAGGAGAGTTTGGCGAGAAGAGAAAAAACTC AATCCTCAATCCCATCAATTCTATTAGAAAGTTTAGCATCGTCCAAAAAACACCATT GCAGATGAATGGGATTGAGGAGGACAGTGATGAGCCTTTGGAACGAAGACTGTCCC TGGTACCCGATAGCGAACAGGGTGAGGCCATCCTTCCTAGGATCTCGGTCATAAGTA CAGGGCCCACACTGCAGGCCAGGCGACGTCAAAGTGTCCTCAATCTTATGACGCAC AGTGTGAATCAGGGGCAGAACATCCATCGTAAGACGACAGCTTCAACTCGAAAGGT CAGTCTAGCTCCACAAGCCAATCTTACAGAGCTGGACATTTATTCCCGCCGCCTCAG TCAGGAGACCGGATTGGAAATATCAGAGGAAATTAATGAAGAGGATCTGAAGGAAT GCTTCTTTGATGACATGGAATCGATCCCCGCTGTTACTACCTGGAACACATATCTGA GATATATTACCGTCCATAAGAGCTTAATCTTTGTACTGATATGGTGCTTGGTGATTTT CCTGGCAGAGGTTGCGGCGAGTTIGGTCGTIGCTATGGCTCCTTGGAAACACTCCCCT GCAGGATAAGGGGAACTCCACTCATAGCAGGAATAACAGCTATGCCGTGATCATCA CCTCTACCTCCTCTTATTACGTGTTTTACATATACGTCGGTGTTGCGGATACCCTGTT GGCAATGGGGTTCTTTAGAGGACTACCCCTAGTTCACACCCTGATCACCGTTTCGAA GATCTTGCACCACAAGATGCTTCATAGCGTTCTCCAAGCTCCTATGAGCACCCTTAA TACACTGAAAGCAGGAGGTATCCTTAACCGCTTTTCCAAAGACATCGCTATACTCGA CGATTTGCTCCCATTGACCATCTTCGACTTCATTCAGCTGCTCCTCATTGTGATCGGC GCCATTGCCGTGGTCGCAGTGTTACAGCCATATATTTTCGTAGCCACCGTGCCCGTC ATCGTGGCATTTATCATGCTGCGCGCATATTTCTTACAGACATCTCAGCAACTGAAG CAGCTGGAATCTGAGGGCAGATCTCCTATTTTTACACACCTGGTTACCAGCCTGAAG GGCCTGTGGACCCTGCGTGCTTTCGGTCGCCAACCCTACTTTGAGACTCTCTTCCATA AGGCTCTGAATTTACATACTGCCAATTGGTTCCTATACCTTAGTACCCTTCGGTGGTT CCAGATGCGGATAGAAATGATCTTCGTGATTTTCTTCATCGCAGTCACTTTCATCTCT ATTTTGACGACCGGTGAGGGCGAGGGCAGGGTGGGCATCATTCTGACTTTGGCCATG AACATTATGTCAACACTCCAGTGGGCCGTTAATTCAAGCATTGATGTGGATTCCTTG ATGCGTTCCGTCAGCAGGGTATTTAAATTCATAGACATGCCCACCGAGGGCAAGCCA ACAAAATCTACCAAGCCATACAAAAATGGCCAACTAAGCAAGGTCATGATTATCGA GAATTCTCATGTGAAAAAGGACGACATTTGGCCTTCCGGGGGTCAAATGACTGTAA AGGACCTGACGGCTAAATACACTGAGGGCGGTAATGCTATCTTGGAGAACATCTCTT TCAGCATCTCCCCTGGCCAGAGAGTGGGACTGCTCGGGCGGACAGGCTCCGGAAAG TCTACGCTCCTTTCAGCATTCCTTAGACTTICTGAACACCGAAGGTGAGATTCAGATT GACGGGGTCTCTTGGGACTCCATCACACTTCAGCAATGGAGGAAGGCATTCGGTGTA ATCCCCCAAAAGGTTTTTATCTTCTCCGGAACATTTCGTAAGAATCTGGACCCGTAC GAGCAGTGGTCAGATCAGGAGATCTGGAAAGTAGCAGACGAGGTCGGGCTACGGA GCGTTATTGAACAGTTTCCTGGCAAACTGGACTTCGTTTTGGTGGACGGAGGCTGTG TGCTGAGTCACGGCCATAAACAACTGATGTGCTTAGCTAGGTCTGTTCTCAGCAAGG CAAAGATTTTACTGCTGGATGAACCAAGCGCCCACCTTGATCCAGTGACATATCAAA TCATCAGAAGAACTCTTAAACAGGCGTTCGCCGACTGCACAGTGATCCTGTGTGAGC ACAGAATAGAAGCCATGCTGGAATGTCAACAGTTTCTCGTGATTGAGGAGAACAAG GTGCGCCAGTACGATAGCATCCAGAAGTTACTCAATGAAAGGTCACTCTTCAGGCA GGCCATCTCACCCAGCGACCGCGTTAAGCTGTTTCCACACCGAAACAGTTCCAAGTG CAAAAGTAAGCCACAGATTGCTGCACTGAAGGAAGAGACAGAAGAAGAAGTTCAG GACACTCGGCTCTGA (SEQ ID NO: 38) ATGCAGAGGAGCCCACTGGAGAAAGCCTCCGTGGTGAGTAAACTCTITTITTAGTTGG ACCAGACCCATCCTGCGAAAAGGATACAGGCAGCGCCTCGAGTTGTCAGATATCTA CCAGATTCCTTCTGTGGACTCAGCTGACAATTTGAGTGAGAAGCTGGAGCGGGAGTG GGATAGAGAGCTGGCGAGCAAAAAAAACCCCAAGCTTATCAATGCTCTGCGCCGCT GCTTTTTCTGGAGGTTCATGTTTTATGGGATCTTCCTGTACCTGGGGGAGGTCACCAA AGCTGTTCAGCCGCTCCTTCTTGGCCGCATCATCGCCAGCTATGACCCTGATAATAA AGAAGAAAGGTCTATTGCTATTTATCTGGGAATTGGCCTCTGCTTGCTCTTCATCGTC CGCACCCTTCTGCTGCACCCTGCCATTTTTGGCCTTCACCACATCGGCATGCAAATG AGAATTGCCATGTTCTCCCTCATTTACAAAAAGACCCTGAAACTTTCCTCAAGAGTG TTAGATAAAATATCCATTGGTCAGCTGGTCAGCCTGCTGTCCAACAATCTTAACAAA TTTGATGAAGGCTTGGCGCTGGCCCACTTCGTGTGGATTGCACCTCTGCAGGTGGCC CTGTTGATGGGACTTATATGGGAGCTGCTTCAAGCCTCTGCTTTCTGTGGGCTGGGCT TTTTGATTGTACTGGCACTTTTTCAGGCTGGGCTCGGAAGAATGATGATGAAATACA GAGATCAGCGGGCCGGGAAGATATCAGAGCGACTTGTGATCACCAGTGAAATGATT GAAAATATTCAGAGCGTGAAAGCCTACTGCTGGGAAGAAGCCATGGAGAAGATGAT TGAGAACCTGAGGCAGACAGAGCTCAAGCTCACTCGGAAGGCTGCTTATGTTCGCT ATTTCAACAGCAGCGCCTTCTTCTTCAGTGGCTTCTTITGTTGTCTTCCTGTCTGTTCTG CCATATGCACTGATAAAAGGCATTATTTTACGAAAGATCTTCACCACCATCAGTTTT TGCATCGTTCTCAGGATGGCCGTCACAAGACAGTTCCCCTGGGCTGTGCAGACCTGG TACGATTCCTTGGGGGCCATCAACAAGATTCAAGATTTCTTGCAAAAACAAGAATAT AAAACTTTAGAATACAACCTCACCACCACTGAAGTGGTCATGGAAAATGTGACAGC CTTTTGGGAGGAGGGTTTTGGAGAATTGTTCGAGAAGGCAAAGCAGAATAACAACA ACAGGAAGACGAGCAATGGGGACGACTCTCTCTTCTTCAGCAACTITTCACTGCTCG GGACCCCTGTGTTGAAAGATATAAACTTCAAGATCGAGAGGGGCCAGCTCTTGGCT GTGGCAGGCTCCACTGGAGCTGGTAAAACATCTCTTCTCATGGTGATCATGGGGGAA CTGGAGCCTTCCGAAGGAAAAATCAAGCACAGTGGGAGAATCTCATTCTGCAGCCA GTITTTCCTGGATCATGCCCGGCACCATTAAGGAAAACATCATATTITGGAGTGTICCTA TGATGAGTACCGCTACCGGTCAGTCATCAAAGCCTGTCAGTTGGAGGAGGACATCTC CAAGTTTGCAGAGAAAGACAACATTGTGCTTGGAGAGGGGGGTATCACTCTTTCTGG AGGACAAAGAGCCAGGATCTCTTTGGCCCGGGCAGTCTACAAGGATGCAGACCTCT ACTTGTTGGACAGTCCCTTCGGCTACCTCGACGTGCTGACTGAAAAAGAAATTTITG AAAGCTGTGTGTGCAAACTGATGGCAAACAAGACCAGGATTCTTGTCACCAGCAAG ATGGAACATCTGAAGAAAGCGGACAAAATTCTGATTCTGCATGAAGGGAGCTCCTA CTTCTATGGAACATTTAGCGAGCTTCAGAACCTACAGCCAGACTTCTCCTCCAAATT AATGGGCTGTGACTCCTTCGACCAGTTCTCTGCAGAAAGAAGAAACTCTATACTCAC AGAGACCCTCCACCGCTTCTCCCTTGAGGGAGATGCCCCAGTTTCTTGGACAGAAAC CAAGAAGCAGTCCTTTAAGCAGACTGGCGAGTTTGGTGAAAAGAGGAAAAATTCAA TTCTCAATCCAATTAACAGTATTCGCAAGTTCAGCATTGTCCAGAAGACACCCCTCC AGATGAATGGCATCGAAGAAGATAGTGACGAGCCGCTGGAGAGACGGCTGAGTCTG GTGCCAGATTCAGAACAGGGGGAGGCCATCCTGCCCCGGATCAGCGTCATTTCCAC AGGCCCCACATTACAAGCACGGCGCCGGCAGAGTGTTTTAAATCTCATGACCCATTC AGTGAACCAGGGCCAAAATATCCACAGGAAGACTACAGCTTCTACCCGGAAAGTGT CTCTGGCCCCTCAGGCCAATCTGACCGAGCTGGACATCTACAGCAGGAGGCTCTCCC AGGAAACAGGGCTGGAAATATCTGAAGAGATTAATGAAGAGGATCTTAAAGAGTGC TTCTTTGATGACATGGAGAGCATCCCCGCGGTGACCACATGGAACACCTACCTTAGA TATATTACTGTCCACAAGAGCCTCATATTTGTCCTCATCTGGTGCCTGGTTATTTTCC TCGCTGAGGTGGCGGCCAGTCTTGTTGTGCTCTGGCTGCTGGGCAACACTCCTCTCC AGGACAAGGGCAATAGTACTCACAGCAGAAATAATTCTTATGCCGTCATCATTACA AGCACCTCCAGCTACTACGTGTTCTACATCTATGTGGGCGTGGCTGACACCCTCCTG GCCATGGGTTTCTTCCGGGGCCTGCCTTTGGTGCACACCCTCATCACAGTGTCAAAA ATTCTGCACCATAAAATGCTTCATTCTGTCCTGCAGGCACCCATGAGCACTTTGAAC ACATTGAAGGCTGGCGGCATCCTCAACAGATTTTCTAAAGATATTGCTATCCTGGAT GATCTCCTCCCCCTGACAATCTTTGACTTTATCCAGCTTCTGCTGATCGTGATTGGAG CCATAGCAGTGGTTGCTGTCCTGCAGCCCTACATTTITGTGGCCACCGTGCCCGTGAT TGTTGCCTTTATTATGCTCAGAGCTTACTTCCTGCAAACTTCTCAACAGCTCAAACAG CTAGAATCTGAGGGCCGGAGCCCCATTTTTACCCACCTGGTGACTTCCCTGAAGGGA CTGTGGACTCTGAGAGCATTCGGGCGACAGCCTTACTTTGAGACACTGTTCCACAAG GCCCTGAACTTIGCACACTGCCAACTGGTITCTITTACCTGAGCACACTCCGCTGGTTCC AGATGCGGATAGAGATGATCTTCGTCATCTTTTTTATAGCTGTAACCTTCATTTICTAT CCTTACAACAGGAGAAGGAGAGGGCAGGGTGGGAATCATCCTCACGCTGGCTATGA ACATAATGTCCACCTTGCAGTGGGCCGTGAATTCCAGTATAGATGTGGATTCTCTAA TGAGGAGTGTCTCCCGGGTGTTTAAATTCATTGATATGCCTACTGAGGGGAAACCCA CCAAGTCAACAAAACCTTATAAGAATGGACAGCTGAGCAAGGTGATGATAATTGAG AACAGCCACGTGAAGAAGGATGACATTTGGCCCAGCGGGGGCCAGATGACTGTGAA GGACCTGACGGCCAAGTACACCGAAGGTGGAAATGCCATTTTGGAAAACATCAGCT TCTCAATCTCTCCTGGGCAGAGAGTTGGATTGCTGGGTCGCACGGGCAGCGGCAAAT CAACCCTGCTCAGTGCCTTCCTTCGGCTCCTGAATACAGAAGGCGAAATCCAAATTG ACGGGGTGAGCTGGGACAGCATCACCCTGCAGCAGTGGAGAAAAGCATTTGGGGTC ATTCCACAGAAAGTTTTCATCTTCTCTGGCACTTTCAGAAAGAACCTGGACCCCTAT GAGCAGTGGAGCGACCAGGAGATCTGGAAGGTTGCAGATGAAGTTGGCCTGCGGAG TGTGATAGAACAATTTCCTGGCAAGCTGGATTTTGTGCTGGTAGATGGAGGCTGCGT GCTGTCCCACGGCCACAAACAGCTGATGTGCCTCGCCCGCTCCGTTCTTTCAAAGGC CAAAATCTTGCTTTTGGATGAGCCCAGTGCTCACCTCGACCCAGTGACCTATCAGAT AATCCGCAGGACCTTAAAGCAAGCTTITGCCGACTGCACCGTCATACTGTGTGAGCA CCGGATTGAAGCAATGCTGGAATGCCAGCAGTTTCTGGTGATCGAGGAGAATAAGG TCCGGCAGTACGACAGCATCCAGAAGTTGTTGAATGAGCGCAGCCTTITCCGCCAGG CCATCTCCCCATCTGACAGAGTCAAGCTGTTTCCACATAGGAACTCCTCTAAGTGCA AGTCCAAGCCCCAGATCGCTGCCCTCAAGGAGGAAACTGAGGAAGAGGTGCAGGAT ACCCGCCTGTGA (SEQ ID NO: 39) ATGCAACGGAGTCCTCTGGAAAAAGCCTCTGTCGTATCTAAGCTTTTCTTCAGTTGG ACACGCCCGATTTTGAGAAAGGGTTATCGGCAACGCTTGGAACTTAGTGACATCTAC CAAATTCCAAGTGTAGACTCAGCCGATAACTTGAGCGAAAAGCTCGAACGAGAGTG GGATCGAGAACTGGCTAGCAAAAAAAATCCCAAACTCATAAATGCCCTGCGACGCT GTTTCTTTTGGCGATTTATGTTTTACGGTATTTTCCTTTATTTGGGTGAGGTCACGAA GGCTGTACAGCCACTGCTGCTGGGTCGCATCATTGCCTCTTACGACCCTGACAACAA AGAGGAGCGGTCAATAGCTATCTACCTTGGTATAGGACTTTGCTTGCTCTTCATAGT CCGCACGTTGCTTCTCCACCCTGCTATATTTGGTCTCCATCACATTGGGATGCAAATG CGGATCGCGATGTITCAGTCTTATATATAAAAAGACTCTTAAACTTTCCAGCCGGGTT CTGGATAAGATCTCTATTGGTCAACTGGTATCTCTTTTGTCTAACAACCTGAATAAGT TCGACGAGGGCCTTGCATTGGCCCATTTTGTATGGATTGCCCCTTTGCAAGTCGCCCT CCTGATGGGATTGATCTGGGAACTCCTGCAAGCTAGTGCTTTTTGCGGATTGGGATT CCTCATAGTCCTTGCGCTCTTTCAGGCGGGACTTGGACGCATGATGATGAAGTATCG CGACCAACGAGCTGGCAAGATCAGTGAACGGCTTGTAATAACCAGTGAAATGATAG AGAACATCCAGAGCGTAAAAGCTTACTGTTGGGAAGAAGCGATGGAAAAGATGATT GAGAACCTTCGCCAGACAGAACTTAAACTTACACGAAAGGCCGCTTATGTCCGGTA CTTCAACTCTTCAGCATTTTTTTITTAGTGGCTTCTTTGTAGTGTTCCTGTCCGTCCTTC CGTATGCACTTATCAAGGGTATAATACTTAGGAAAATCTTCACAACAATCAGTTTTT GCATAGTCCTTCGCATGGCAGTAACTCGCCAATTTCCCTGGGCAGTTCAGACGTGGT ACGACTCACTTGGCGCAATTAACAAAATTCAAGATTTCCTCCAAAAGCAAGAGTATA AAACCTTGGAATACAACCTTACCACCACAGAAGTTGTAATGGAAAATGTCACAGCC TTCTGGGAGGAAGGTTTCGGCGAACTTTTTGAGAAGGCGAAGCAAAATAACAATAA TCGGAAAACATCAAACGGTGACGATTCACTGTTCTTTTCTAACTTTAGCCTTCTTGGG ACGCCCGTCCTGAAGGACATAAACTTTAAGATTGAACGGGGTCAACTTCTCGCGGTC GCAGGGAGTACTGGAGCGGGGAAAACGAGCCTGCTGATGGTGATAATGGGGGAGTT GGAGCCCTCAGAAGGCAAGATCAAGCATAGTGGTAGAATTAGCTTCTGCAGTCAAT TTAGTTGGATTATGCCGGGCACGATCAAAGAAAATATAATCTTTGGGGTATCCTACG ATGAATACAGGTACCGATCAGTGATAAAAGCGTGCCAGCTTGAAGAAGACATTTCA AAGTTTGCTGAGAAGGATAATATCGTACTTGGAGAAGGAGGTATCACCCTGTCTGG GGGTCAACGAGCGAGGATCTCCCTGGCACGCGCCGTCTACAAGGACGCGGACCTCT ATCTGTTGGATTCACCGTTCGGATATTTGGACGTGCTTACGGAGAAAGAAATATITG AGAGCTGTGTTTGCAAGCTCATGGCAAATAAAACCAGAATATTGGTTACAAGCAAG ATGGAGCATCTTAAGAAAGCAGATAAAATCCTGATATTGCACGAGGGCTCTTCATAC TTCTACGGGACGTTTTCTGAGTTGCAGAACCTCCAGCCGGATTTCAGCTCTAAGCTG ATGGGCTGTGATTCCTTTGATCAGTTTAGTGCGGAAAGACGAAACAGTATACTCACC GAAACACTGCACAGGTTCTCTCTGGAGGGCGACGCCCCGGTTTCCTGGACAGAGAC GAAGAAGCAGTCCTTCAAACAGACAGGCGAGTTTGGGGAGAAAAGGAAAAATAGC ATACTCAACCCGATTAACAGCATTCGCAAGTTCAGTATAGTACAAAAGACCCCGTTG CAGATGAACGGTATAGAGGAAGATTCTGATGAGCCACTGGAAAGACGGCTTTICTCT CGTTCCGGACAGTGAACAGGGAGAGGCAATACTGCCTCGGATCAGCGTTATCTCTAC AGGACCTACTTTGCAAGCTCGGCGCCGACAGTCAGTCTTGAATCTTATGACTCATAG TGTTAATCAAGGCCAGAATATCCATCGCAAGACCACCGCAAGTACAAGGAAAGTGA GCTTGGCACCTCAAGCAAACCTTACTGAACTTGATATCTACTCACGGCGACTTTCAC AGGAGACCGGACTTGAAATTAGTGAAGAAATTAACGAGGAGGACCTCAAGGAGTGC TTCTTCGATGACATGGAATCAATCCCCGCAGTCACAACCTGGAACACTTATCTGAGG TATATAACAGTTCACAAGAGCCTCATTTTTGTACTTATTTGGTGTTTGGTAATTTTCC TGGCGGAGGTTGCTGCTTCTTTGGTCGTCCTTTGGCTCCTCGGGAATACACCGCTCCA AGACAAAGGCAACTCTACCCATAGTAGGAACAATTCATATGCAGTGATTATAACCA GTACATCATCTTATTACGTTTTCTATATTTATGTCGGGGTAGCTGACACGCTGTTGGC GATGGGCTTCTTTAGGGGCCTCCCCTTGGTACACACCCTTATCACGGTGAGTAAAAT CCTGCATCACAAAATGCTTCATTCTGTACTCCAAGCGCCGATGAGTACGCTTAATAC GCTGAAAGCAGGAGGGATACTGAATCGGTTCAGCAAGGACATCGCCATTCTGGATG ACCTGCTTCCATTGACAATATTTGATTTCATTCAGCTCCTTCTCATAGTTATTGGAGC CATAGCGGTGGTGGCTGTGCTTCAGCCTTATATATTCGTTGCCACAGTTCCCGTTATA GTGGCATTTATAATGCTCAGGGCCTACTTTCTCCAGACTTCCCAGCAGTTGAAGCAA CTCGAATCAGAAGGAAGGTCACCTATTTTCACACATCTTGTGACTTCCTTGAAGGGC TTGTGGACGCTGCGGGCCTTCGGAAGACAACCATATTTTGAAACTCTCTTCCACAAA GCTTTGAATCTTCATACTGCGAACTGGTTCCTGTATTTGAGTACTTTGCGCTGGTTCC AGATGAGGATAGAAATGATATTCGTTATCTTCTTTATCGCGGTTACGTTCATAAGTA TCCTCACTACGGGGGAGGGTGAGGGTAGAGTGGGCATAATACTGACCCTCGCCATG AACATTATGTCCACCCTGCAGTGGGCGGTAAACAGCAGCATAGATGTGGATTCTTITG ATGCGCAGTGTGAGCAGGGTTTTTAAGTTTATCGATATGCCGACGGAAGGAAAGCC CACTAAAAGCACGAAACCCTATAAAAATGGACAGCTTAGCAAAGTAATGATAATCG AGAATAGCCATGTGAAAAAGGATGACATATGGCCTTCCGGAGGCCAAATGACTGIT AAAGATCTGACCGCTAAATATACCGAGGGCGGCAACGCAATACTCGAAAACATAAG CTTTTCCATAAGCCCCGGCCAACGCGTGGGTCTTCTGGGGAGGACTGGCTCCGGAAA ATCAACGTTGCTTAGCGCGTTTTTGCGGCTCCTTAACACTGAAGGTGAGATCCAAAT AGATGGCGTTAGTTGGGACTCTATAACACTGCAACAATGGCGGAAAGCTTTCGGCGT CATACCTCAGAAGGTGTTCATCTTTAGCGGAACGTTCAGGAAGAACTTGGATCCCTA CGAACAATGGAGTGATCAAGAAATATGGAAAGTGGCAGATGAGGTAGGCTTGCGCA GTGTCATTGAACAATTCCCAGGGAAACTCGACTTTGTACTGGTGGACGGCGGTTGCG TCTTGTCACACGGGCACAAACAGTTGATGTGTTTGGCCCGCAGTGTTTTGTCTAAGG CGAAGATTCTGTTGCTCGACGAACCGAGTGCTCATCTTGATCCCGTCACCTACCAAA TCATCAGAAGGACGTTGAAGCAAGCTTTCGCCGACTGCACTGTAATCCTTTGTGAGC ATAGGATCGAAGCAATGCTCGAGTGCCAACAGTTCTTGGTTATAGAGGAGAATAAG GTTCGGCAATACGACTCAATACAGAAACTGCTTAATGAGCGGTCACTCTTTCGACAA GCTATCTCTCCTAGTGACAGGGTAAAGCTTTTTCCTCATCGGAATTCCAGCAAGTGT AAGAGTAAACCACAGATCGCCGCCCTTAAAGAGGAGACCGAAGAAGAGGTGCAGG ATACGAGACTTTAG (SEQ ID NO: 40) Synthesis of mRNA

[0128] mRNAs according to the present invention may be synthesized according to any of a variety of known methods. For example, mRNAs according to the present invention may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g, T3, T7, or SP6 RNA polymerase), DNAse I, pyrophosphatase, and / or RNAse inhibitor. The exact conditions will vary according to the specific application. Exemplary Codon-optintized Human Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) mRNAs Construct design: X~SEQIDNO:1-Y 5'and 3' UTR Sequences: X (5' UTR Sequence) = GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACC GGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCG UGCCAAGAGUGACUCACCGUCCUUGACACG (SEQ ID NO: 4) Y (3' UTR Sequence) = CGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCC ACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAGCU (SEQ 1D NO: 5) OR GGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCA CUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAAGCU (SEQ ID NO: 6)

[0129] An exemplary codon-optimized human CFTR mRNA sequence includes SEQ ID NO: 1 as described in the detailed description section.

[0130] An exemplary full-length codon-optimized human CFTR mRNA sequence is shown below: GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACC GGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCG UGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAACGCUCUCCUCUUGAAAAGG CCUCGGUGGUGUCCAAGCUCUUCUUCUCGUGGACUAGACCCAUCCUGAGAAAGGG GUACAGACAGCGCUUGGAGCUGUCCGAUAUCUAUCAAAUCCCUUCCGUGGACUCC GCGGACAACCUGUCCGAGAAGCUCGAGAGAGAAUGGGACAGAGAACUCGCCUCAA AGAAGAACCCGAAGCUGAUUAAUGCGCUUAGGCGGUGCUUUUUCUGGCGGUUCA UGUUCUACGGCAUCUUCCUCUACCUGGGAGAGGUCACCAAGGCCGUGCAGCCCCU GUUGCUGGGACGGAUUAUUGCCUCCUACGACCCCGACAACAAGGAAGAAAGAAGC AUCGCUAUCUACUUGGGCAUCGGUCUGUGCCUGCUUUUCAUCGUCCGGACCCUCU UGUUGCAUCCUGCUAUUUUCGGCCUGCAUCACAUUGGCAUGCAGAUGAGAAUUG CCAUGUUUUCCCUGAUCUACAAGAAAACUCUGAAGCUCUCGAGCCGCGUGCUUGA CAAGAUUUCCAUCGGCCAGCUCGUGUCCCUGCUCUCCAACAAUCUGAACAAGUUC GACGAGGGCCUCGCCCUGGCCCACUUCGUGUGGAUCGCCCCUCUGCAAGUGGCGC UUCUGAUGGGCCUGAUCUGGGAGCUGCUGCAAGCCUCGGCAUUCUGUGGGCUUG GAUUCCUGAUCGUGCUGGCACUGUUCCAGGCCGGACUGGGGCGGAUGAUGAUGA AGUACAGGGACCAGAGAGCCGGAAAGAUUUCCGAACGGCUGGUGAUCACUUCGG AAAUGAUCGAAAACAUCCAGUCAGUGAAGGCCUACUGCUGGGAAGAGGCCAUGG AAAAGAUGAUUGAAAACCUCCGGCAAACCGAGCUGAAGCUGACCCGCAAGGCCGC UUACGUGCGCUAUUUCAACUCGUCCGCUUUCUUCUUCUCCGGGUUCUUCGUGGUG UUUCUCUCCGUGCUCCCCUACGCCCUGAUUAAGGGAAUCAUCCUCAGGAAGAUCU UCACCACCAUUUCCUUCUGUAUCGUGCUCCGCAUGGCCGUGACCCGGCAGUUCCC AUGGGCCGUGCAGACUUGGUACGACUCCCUGGGAGCCAUUAACAAGAUCCAGGAC UUCCUUCAAAAGCAGGAGUACAAGACCCUCGAGUACAACCUGACUACUACCGAGG UCGUGAUGGAAAACGUCACCGCCUUUUGGGAGGAGGGAUUUGGCGAACUGUUCG AGAAGGCCAAGCAGAACAACAACAACCGCAAGACCUCGAACGGUGACGACUCCCU CUUCUUUUCAAACUUCAGCCUGCUCGGGACGCCCGUGCUGAAGGACAUUAACUUC AAGAUCGAAAGAGGACAGCUCCUGGCGGUGGCCGGAUCGACCGGAGCCGGAAAG ACUUCCCUGCUGAUGGUGAUCAUGGGAGAGCUUGAACCUAGCGAGGGAAAGAUC AAGCACUCCGGCCGCAUCAGCUUCUGUAGCCAGUUUUCCUGGAUCAUGCCCGGAA CCAUUAAGGAAAACAUCAUCUUCGGCGUGUCCUACGAUGAAUACCGCUACCGGUC CGUGAUCAAAGCCUGCCAGCUGGAAGAGGAUAUUUCAAAGUUCGCGGAGAAAGA UAACAUCGUGCUGGGCGAAGGGGGUAUUACCUUGUCGGGGGGCCAGCGGGCUAG AAUCUCGCUGGCCAGAGCCGUGUAUAAGGACGCCGACCUGUAUCUCCUGGACUCC CCCUUCGGAUACCUGGACGUCCUGACCGAAAAGGAGAUCUUCGAAUCGUGCGUGU GCAAGCUGAUGGCUAACAAGACUCGCAUCCUCGUGACCUCCAAAAUGGAGCACCU GAAGAAGGCAGACAAGAUUCUGAUUCUGCAUGAGGGGUCCUCCUACUUUUACGG CACCUUCUCGGAGUUGCAGAACUUGCAGCCCGACUUCUCAUCGAAGCUGAUGGGU UGCGACAGCUUCGACCAGUUCUCCGCCGAAAGAAGGAACUCGAUCCUGACGGAAA CCUUGCACCGCUUCUCUUUGGAAGGCGACGCCCCUGUGUCAUGGACCGAGACUAA GAAGCAGAGCUUCAAGCAGACCGGGGAAUUCGGCGAAAAGAGGAAGAACAGCAU CUUGAACCCCAUUAACUCCAUCCGCAAGUUCUCAAUCGUGCAAAAGACGCCACUG CAGAUGAACGGCAUUGAGGAGGACUCCGACGAACCCCUUGAGAGGCGCCUGUCCC UGGUGCCGGACAGCGAGCAGGGAGAAGCCAUCCUGCCUCGGAUUUCCGUGAUCUC CACUGGUCCGACGCUCCAAGCCCGGCGGCGGCAGUCCGUGCUGAACCUGAUGACC CACAGCGUGAACCAGGGCCAAAACAUUCACCGCAAGACUACCGCAUCCACCCGGA AAGUGUCCCUGGCACCUCAAGCGAAUCUUACCGAGCUCGACAUCUACUCCCGGAG ACUGUCGCAGGAAACCGGGCUCGAAAUUUCCGAAGAAAUCAACGAGGAGGAUCU GAAAGAGUGCUUCUUCGACGAUAUGGAGUCGAUACCCGCCGUGACGACUUGGAA CACUUAUCUGCGGUACAUCACUGUGCACAAGUCAUUGAUCUUCGUGCUGAUUUG GUGCCUGGUGAUUUUCCUGGCCGAGGUCGCGGCCUCACUGGUGGUGCUCUGGCUG UUGGGAAACACGCCUCUGCAAGACAAGGGAAACUCCACGCACUCGAGAAACAACA GCUAUGCCGUGAUUAUCACUUCCACCUCCUCUUAUUACGUGUUCUACAUCUACGU CGGAGUGGCGGAUACCCUGCUCGCGAUGGGUUUCUUCAGAGGACUGCCGCUGGUC CACACCUUGAUCACCGUCAGCAAGAUUCUUCACCACAAGAUGUUGCAUAGCGUGC UGCAGGCCCCCAUGUCCACCCUCAACACUCUGAAGGCCGGAGGCAUUCUGAACAG AUUCUCCAAGGACAUCGCUAUCCUGGACGAUCUCCUGCCGCUUACCAUCUUUGAC UUCAUCCAGCUGCUGCUGAUCGUGAUUGGAGCAAUCGCAGUGGUGGCGGUGCUG CAGCCUUACAUUUUCGUGGCCACUGUGCCGGUCAUUGUGGCGUUCAUCAUGCUGC GGGCCUACUUCCUCCAAACCAGCCAGCAGCUGAAGCAACUGGAAUCCGAGGGACG AUCCCCCAUCUUCACUCACCUUGUGACGUCGUUGAAGGGACUGUGGACCCUCCGG GCUUUCGGACGGCAGCCCUACUUCGAAACCCUCUUCCACAAGGCCCUGAACCUCC ACACCGCCAAUUGGUUCCUGUACCUGUCCACCCUGCGGUGGUUCCAGAUGCGCAU CGAGAUGAUUUUCGUCAUCUUCUUCAUCGCGGUCACAUUCAUCAGCAUCCUGACU ACCGGAGAGGGAGAGGGACGGGUCGGAAUAAUCCUGACCCUCGCCAUGAACAUU AUGAGCACCCUGCAGUGGGCAGUGAACAGCUCGAUCGACGUGGACAGCCUGAUGC GAAGCGUCAGCCGCGUGUUCAAGUUCAUCGACAUGCCUACUGAGGGAAAACCCAC UAAGUCCACUAAGCCCUACAAAAAUGGCCAGCUGAGCAAGGUCAUGAUCAUCGAA AACUCCCACGUGAAGAAGGACGAUAUUUGGCCCUCCGGAGGUCAAAUGACCGUGA AGGACCUGACCGCAAAGUACACCGAGGGAGGAAACGCCAUUCUCGAAAACAUCAG CUUCUCCAUUUCGCCGGGACAGCGGGUCGGCCUUCUCGGGCGGACCGGUUCCGGG AAGUCAACUCUGCUGUCGGCUUUCCUCCGGCUGCUGAAUACCGAGGGGGAAAUCC AAAUUGACGGCGUGUCUUGGGAUUCCAUUACUCUGCAGCAGUGGCGGAAGGCCU UCGGCGUGAUCCCCCAGAAGGUGUUCAUCUUCUCGGGUACCUUCCGGAAGAACCU GGAUCCUUACGAGCAGUGGAGCGACCAAGAAAUCUGGAAGGUCGCCGACGAGGU CGGCCUGCGCUCCGUGAUUGAACAAUUUCCUGGAAAGCUGGACUUCGUGCUCGUC GACGGGGGAUGUGUCCUGUCGCACGGACAUAAGCAGCUCAUGUGCCUCGCACGGU CCGUGCUCUCCAAGGCCAAGAUUCUGCUGCUGGACGAACCUUCGGCCCACCUGGA UCCGGUCACCUACCAGAUCAUCAGGAGGACCCUGAAGCAGGCCUUUGCCGAUUGC ACCGUGAUUCUCUGCGAGCACCGCAUCGAGGCCAUGCUGGAGUGCCAGCAGUUCC UGGUCAUCGAGGAGAACAAGGUCCGCCAAUACGACUCCAUUCAAAAGCUCCUCAA CGAGCGGUCGCUGUUCAGACAAGCUAUUUCACCGUCCGAUAGAGUGAAGCUCUUC CCGCAUCGGAACAGCUCAAAGUGCAAAUCGAAGCCGCAGAUCGCAGCCUUGAAGG AAGAGACUGAGGAAGAGGUGCAGGACACCCGGCUUUAACGGGUGGCAUCCCUGU GACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCA GCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAGCU (SEQ ID NO: 7)

[0131] In another example, a full length codon-optimized human CFTR mRNA sequence 1s shown below: GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACC GGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCG UGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAACGCUCUCCUCUUGAAAAGG CCUCGGUGGUGUCCAAGCUCUUCUUCUCGUGGACUAGACCCAUCCUGAGAAAGGG GUACAGACAGCGCUUGGAGCUGUCCGAUAUCUAUCAAAUCCCUUCCGUGGACUCC GCGGACAACCUGUCCGAGAAGCUCGAGAGAGAAUGGGACAGAGAACUCGCCUCAA AGAAGAACCCGAAGCUGAUUAAUGCGCUUAGGCGGUGCUUUUUCUGGCGGUUCA UGUUCUACGGCAUCUUCCUCUACCUGGGAGAGGUCACCAAGGCCGUGCAGCCCCU GUUGCUGGGACGGAUUAUUGCCUCCUACGACCCCGACAACAAGGAAGAAAGAAGC AUCGCUAUCUACUUGGGCAUCGGUCUGUGCCUGCUUUUCAUCGUCCGGACCCUCU UGUUGCAUCCUGCUAUUUUCGGCCUGCAUCACAUUGGCAUGCAGAUGAGAAUUG CCAUGUUUUCCCUGAUCUACAAGAAAACUCUGAAGCUCUCGAGCCGCGUGCUUGA CAAGAUUUCCAUCGGCCAGCUCGUGUCCCUGCUCUCCAACAAUCUGAACAAGUUC GACGAGGGCCUCGCCCUGGCCCACUUCGUGUGGAUCGCCCCUCUGCAAGUGGCGC UUCUGAUGGGCCUGAUCUGGGAGCUGCUGCAAGCCUCGGCAUUCUGUGGGCUUG GAUUCCUGAUCGUGCUGGCACUGUUCCAGGCCGGACUGGGGCGGAUGAUGAUGA AGUACAGGGACCAGAGAGCCGGAAAGAUUUCCGAACGGCUGGUGAUCACUUCGG AAAUGAUCGAAAACAUCCAGUCAGUGAAGGCCUACUGCUGGGAAGAGGCCAUGG AAAAGAUGAUUGAAAACCUCCGGCAAACCGAGCUGAAGCUGACCCGCAAGGCCGC UUACGUGCGCUAUUUCAACUCGUCCGCUUUCUUCUUCUCCGGGUUCUUCGUGGUG UUUCUCUCCGUGCUCCCCUACGCCCUGAUUAAGGGAAUCAUCCUCAGGAAGAUCU UCACCACCAUUUCCUUCUGUAUCGUGCUCCGCAUGGCCGUGACCCGGCAGUUCCC AUGGGCCGUGCAGACUUGGUACGACUCCCUGGGAGCCAUUAACAAGAUCCAGGAC UUCCUUCAAAAGCAGGAGUACAAGACCCUCGAGUACAACCUGACUACUACCGAGG UCGUGAUGGAAAACGUCACCGCCUUUUGGGAGGAGGGAUUUGGCGAACUGUUCG AGAAGGCCAAGCAGAACAACAACAACCGCAAGACCUCGAACGGUGACGACUCCCU CUUCUUUUCAAACUUCAGCCUGCUCGGGACGCCCGUGCUGAAGGACAUUAACUUC AAGAUCGAAAGAGGACAGCUCCUGGCGGUGGCCGGAUCGACCGGAGCCGGAAAG ACUUCCCUGCUGAUGGUGAUCAUGGGAGAGCUUGAACCUAGCGAGGGAAAGAUC AAGCACUCCGGCCGCAUCAGCUUCUGUAGCCAGUUUUCCUGGAUCAUGCCCGGAA CCAUUAAGGAAAACAUCAUCUUCGGCGUGUCCUACGAUGAAUACCGCUACCGGUC CGUGAUCAAAGCCUGCCAGCUGGAAGAGGAUAUUUCAAAGUUCGCGGAGAAAGA UAACAUCGUGCUGGGCGAAGGGGGUAUUACCUUGUCGGGGGGCCAGCGGGCUAG AAUCUCGCUGGCCAGAGCCGUGUAUAAGGACGCCGACCUGUAUCUCCUGGACUCC CCCUUCGGAUACCUGGACGUCCUGACCGAAAAGGAGAUCUUCGAAUCGUGCGUGU GCAAGCUGAUGGCUAACAAGACUCGCAUCCUCGUGACCUCCAAAAUGGAGCACCU GAAGAAGGCAGACAAGAUUCUGAUUCUGCAUGAGGGGUCCUCCUACUUUUACGG CACCUUCUCGGAGUUGCAGAACUUGCAGCCCGACUUCUCAUCGAAGCUGAUGGGU UGCGACAGCUUCGACCAGUUCUCCGCCGAAAGAAGGAACUCGAUCCUGACGGAAA CCUUGCACCGCUUCUCUUUGGAAGGCGACGCCCCUGUGUCAUGGACCGAGACUAA GAAGCAGAGCUUCAAGCAGACCGGGGAAUUCGGCGAAAAGAGGAAGAACAGCAU CUUGAACCCCAUUAACUCCAUCCGCAAGUUCUCAAUCGUGCAAAAGACGCCACUG CAGAUGAACGGCAUUGAGGAGGACUCCGACGAACCCCUUGAGAGGCGCCUGUCCC UGGUGCCGGACAGCGAGCAGGGAGAAGCCAUCCUGCCUCGGAUUUCCGUGAUCUC CACUGGUCCGACGCUCCAAGCCCGGCGGCGGCAGUCCGUGCUGAACCUGAUGACC CACAGCGUGAACCAGGGCCAAAACAUUCACCGCAAGACUACCGCAUCCACCCGGA AAGUGUCCCUGGCACCUCAAGCGAAUCUUACCGAGCUCGACAUCUACUCCCGGAG ACUGUCGCAGGAAACCGGGCUCGAAAUUUCCGAAGAAAUCAACGAGGAGGAUCU GAAAGAGUGCUUCUUCGACGAUAUGGAGUCGAUACCCGCCGUGACGACUUGGAA CACUUAUCUGCGGUACAUCACUGUGCACAAGUCAUUGAUCUUCGUGCUGAUUUG GUGCCUGGUGAUUUUCCUGGCCGAGGUCGCGGCCUCACUGGUGGUGCUCUGGCUG UUGGGAAACACGCCUCUGCAAGACAAGGGAAACUCCACGCACUCGAGAAACAACA GCUAUGCCGUGAUUAUCACUUCCACCUCCUCUUAUUACGUGUUCUACAUCUACGU CGGAGUGGCGGAUACCCUGCUCGCGAUGGGUUUCUUCAGAGGACUGCCGCUGGUC CACACCUUGAUCACCGUCAGCAAGAUUCUUCACCACAAGAUGUUGCAUAGCGUGC UGCAGGCCCCCAUGUCCACCCUCAACACUCUGAAGGCCGGAGGCAUUCUGAACAG AUUCUCCAAGGACAUCGCUAUCCUGGACGAUCUCCUGCCGCUUACCAUCUUUGAC UUCAUCCAGCUGCUGCUGAUCGUGAUUGGAGCAAUCGCAGUGGUGGCGGUGCUG CAGCCUUACAUUUUCGUGGCCACUGUGCCGGUCAUUGUGGCGUUCAUCAUGCUGC GGGCCUACUUCCUCCAAACCAGCCAGCAGCUGAAGCAACUGGAAUCCGAGGGACG AUCCCCCAUCUUCACUCACCUUGUGACGUCGUUGAAGGGACUGUGGACCCUCCGG GCUUUCGGACGGCAGCCCUACUUCGAAACCCUCUUCCACAAGGCCCUGAACCUCC ACACCGCCAAUUGGUUCCUGUACCUGUCCACCCUGCGGUGGUUCCAGAUGCGCAU CGAGAUGAUUUUCGUCAUCUUCUUCAUCGCGGUCACAUUCAUCAGCAUCCUGACU ACCGGAGAGGGAGAGGGACGGGUCGGAAUAAUCCUGACCCUCGCCAUGAACAUU AUGAGCACCCUGCAGUGGGCAGUGAACAGCUCGAUCGACGUGGACAGCCUGAUGC GAAGCGUCAGCCGCGUGUUCAAGUUCAUCGACAUGCCUACUGAGGGAAAACCCAC UAAGUCCACUAAGCCCUACAAAAAUGGCCAGCUGAGCAAGGUCAUGAUCAUCGAA AACUCCCACGUGAAGAAGGACGAUAUUUGGCCCUCCGGAGGUCAAAUGACCGUGA AGGACCUGACCGCAAAGUACACCGAGGGAGGAAACGCCAUUCUCGAAAACAUCAG CUUCUCCAUUUCGCCGGGACAGCGGGUCGGCCUUCUCGGGCGGACCGGUUCCGGG AAGUCAACUCUGCUGUCGGCUUUCCUCCGGCUGCUGAAUACCGAGGGGGAAAUCC AAAUUGACGGCGUGUCUUGGGAUUCCAUUACUCUGCAGCAGUGGCGGAAGGCCU UCGGCGUGAUCCCCCAGAAGGUGUUCAUCUUCUCGGGUACCUUCCGGAAGAACCU GGAUCCUUACGAGCAGUGGAGCGACCAAGAAAUCUGGAAGGUCGCCGACGAGGU CGGCCUGCGCUCCGUGAUUGAACAAUUUCCUGGAAAGCUGGACUUCGUGCUCGUC GACGGGGGAUGUGUCCUGUCGCACGGACAUAAGCAGCUCAUGUGCCUCGCACGGU CCGUGCUCUCCAAGGCCAAGAUUCUGCUGCUGGACGAACCUUCGGCCCACCUGGA UCCGGUCACCUACCAGAUCAUCAGGAGGACCCUGAAGCAGGCCUUUGCCGAUUGC ACCGUGAUUCUCUGCGAGCACCGCAUCGAGGCCAUGCUGGAGUGCCAGCAGUUCC UGGUCAUCGAGGAGAACAAGGUCCGCCAAUACGACUCCAUUCAAAAGCUCCUCAA CGAGCGGUCGCUGUUCAGACAAGCUAUUUCACCGUCCGAUAGAGUGAAGCUCUUC CCGCAUCGGAACAGCUCAAAGUGCAAAUCGAAGCCGCAGAUCGCAGCCUUGAAGG AAGAGACUGAGGAAGAGGUGCAGGACACCCGGCUUUAAGGGUGGCAUCCCUGUG ACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGC CUUGUCCUAAUAAAAUUAAGUUGCAUCAAAGCU (SEQ ID NO: 8)

[0132] In some embodiments, an activity of CFTR proteins is evaluated by an Ussing chamber assay. In some embodiments, duration of activity of CFTR proteins is evaluated by time-course Ussing assays. In some embodiments, protein expression and stability are evaluated by pulse-chase methods. In some embodiments, protein expression and stability are evaluated by surface biotinylation.

[0133] In some embodiments, for the preparation of mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter, for example a T3, T7 or SP6 promoter, for in vitro transcription, followed by desired nucleotide sequence for desired mRNA and a termination signal. Codon optimization

[0134] According to an increasing amount of research, mRNAs contain numerous layers of information that overlap the amino acid code. Traditionally, codon optimization has been used to remove rare codons which were thought to be rate-limiting for protein expression. While fast growing bacteria and yeast both exhibit strong codon bias in highly expressed genes, higher eukaryotes exhibit much less codon bias, making it more difficult to discern codons that may be rate-limiting. Tn addition, it has been found that codon bias per se does not necessarily yield high expression but requires other features.

[0135] For example, rare codons have been implicated in slowing translation and forming pause sites, which may be required for correct protein folding. Therefore, variations in codon usage may provide a mechanism to fine-tune the temporal pattern of elongation and thus increase the time available for a protein to take on its correct confirmation. Codon optimization can interfere with this fine-tuning mechanism, resulting in less efficient protein translation or an increased amount of incorrectly folded proteins. Similarly, codon optimization may disrupt the normal patterns of cognate and wobble tRNA usage, thereby affecting protein structure and function because wobble-dependent slowing of elongation may likewise have been selected as a mechanism for achieving correct protein folding.

[0136] Various methods of performing codon optimization are known in the art, however, each has significant drawbacks and limitations from a computational and / or therapeutic point of view. In particular, known methods of codon optimization often involve, for each amino acid, replacing every codon with the codon having the highest usage for that amino acid, such that the “optimized” sequence contains only one codon encoding each amino acid (so may be referred to as a one-to-one sequence). The increase in expression is not limited to cell cultures of mammalian cells but was also observed i» vivo in a mouse model. It is expected that the observed improvement in expression of the codon-optimised CFTR coding sequence, either wild type or activated CFTR, will result in an improved, more cost-effective mRNA replacement therapy for patients in need thereof, because it does not require the use of modified nucleotides for the preparation of the mRNA and allows treatment with a reduced dose and / or at extended dosing intervals.

[0137] Tn some embodiments, codon-optimized mRNA is produced in accordance with methods known in the art.

[0138] Tn some embodiments, codon-optimized mRNA sequences according to the present invention were further codon-optimized by a new process: the process first generates a list of codon-optimized sequences and then applies three filters to the list. Specifically, it applies a motif screen filter, guanine-cytosine (GC) content analysis filter, and codon adaptation index (CAL) analysis filter to produce an updated list of optimized nucleotide sequences. The updated list no longer includes nucleotide sequences containing features that are expected to interfere with effective transcription and / or translation of the encoded protein antigen. Synthesis of mRNA using SP6 RNA Polymerase

[0139] In some embodiments, CFTR mRNA is produced using SP6 RNA Polymerase. SP6 RNA Polymerase is a DNA-dependent RNA polymerase with high sequence specificity for SP6 promoter sequences. The SP6 polymerase catalyzes the 5'—3' in vitro synthesis of RNA on either single-stranded DNA or double-stranded DNA downstream from its promoter; it incorporates native ribonucleotides and / or modified ribonucleotides and / or labeled ribonucleotides into the polymerized transcript. Examples of such labeled ribonucleotides include biotin-, fluorescein-, digoxigenin-, aminoallyl-, and isotope-labeled nucleotides.

[0140] The sequence for bacteriophage SP6 RNA polymerase was initially described (GenBank: Y00105.1) as having the following amino acid sequence: MQDLHAIQLQLEEEMFNGGIRRFEADQQRQIAAGSESDTAWNRRLLSELIAPMAEGIQA YKEEYEGKKGRAPRALAFLQCVENEVAAYITMKVVMDMLNTDATLQAIAMSVAERIE DQVRFSKLEGHAAKYFEKVKKSLKASRTKS YRHAHNVAVVAEKSVAEKDADFDRWEA WPKETQLQIGTTLLEILEGSVFYNGEPVFMRAMRTYGGKTIY YLQTSESVGQWISAFKE HVAQLSPAYAPCVIPPRPWRTPFNGGFHTEKVASRIRLVKGNREHVRKLTQKQMPKVY KAINALQNTQWQINKDVLAVIEEVIRLDLGYGVPSFKPLIDKENKPANPVPVEFQHLRGR ELKEMLSPEQWQQFINWKGECARLYTAETKRGSKSAAVVRMVGQARKYSAFESIYFVY AMDSRSRVYVQSSTLSPQSNDLGKALLRFTEGRPVNGVEALKWFCINGANLWGWDKK TFDVRVSNVLDEEFQDMCRDIAADPLTFTQWAKADAPYEFLAWCFEYAQYLDLVDEG RADEFRTHLPVHQDGSCSGIQHYSAMLRDEVGAKAVNLKPSDAPQDIYGAVAQVVIKK NALYMDADDATTFTSGSVTLSGTELRAMASAWDSIGITRSLTKKPVMTLPYGSTRLTCR ESVIDYIVDLEEKEAQKAVAEGRTANKVHPFEDDRQDYLTPGAA YNYMTALIWPSISEV VKAPIVAMKMIRQLARFAAKRNEGLMY TLPTGFILEQKIMATEMLRVRTCLMGDIKMS LQVETDIVDEAAMMGAAAPNFVHGHDASHLILTVCELVDKGVTSIAVIHDSFGTHADNT LTLRVALKGQMVAMYIDGNALQKLLEEHEVRWMVDTGIEVPEQGEFDLNEIMDSEY VF A (SEQ ID NO: 9).

[0141] An SP6 RNA polymerase suitable for the present invention can be any enzyme having substantially the same polymerase activity as bacteriophage SP6 RNA polymerase. Thus, in some embodiments, an SP6 RNA polymerase suitable for the present invention may be modified from SEQ ID NO: 9. For example, a suitable SP6 RNA polymerase may contain one or more amino acid substitutions, deletions, or additions. In some embodiments, a suitable SP6 RNA polymerase has an amino acid sequence about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 75%, 70%, 65%, or 60% identical or homologous to SEQ ID NO: 9. In some embodiments, a suitable SP6 RNA polymerase may be a truncated protein (from N-terminus, C-terminus, or internally) but retain the polymerase activity. In some embodiments, a suitable SP6 RNA polymerase is a fusion protein.

[0142] An SP6 RNA polymerase suitable for the invention may be a commercially- available product, e.g., from Aldevron, Ambion, New England Biolabs (NEB), Promega, and Roche. The SP6 may be ordered and / or custom designed from a commercial source or a non- commercial source according to the amino acid sequence of SEQ ID NO: 9 or a variant of SEQ ID NO: 9 as described herein. The SP6 may be a standard-fidelity polymerase or may be a high- fidelity / high-efficiency / high-capacity which has been modified to promote RNA polymerase activities, e.g., mutations in the SP6 RNA polymerase gene or post-translational modifications of the SP6 RNA polymerase itself. Examples of such modified SP6 include SP6 RNA Polymerase- Plus™ from Ambion, HiScribe SP6 from NEB, and RiboMAX™ and Riboprobe® Systems from Promega.

[0143] Tn some embodiments, a suitable SP6 RNA polymerase is a fusion protein. For example, an SP6 RNA polymerase may include one or more tags to promote isolation, purification, or solubility of the enzyme. A suitable tag may be located at the N-terminus, C- terminus, and / or internally. Non-limiting examples of a suitable tag include Calmodulin-binding protein (CBP); Fasciola hepatica 8-kDa antigen (Fh8); FLAG tag peptide; glutathione-S- transferase (GST); Histidine tag (e.g., hexahistidine tag (His6)); maltose-binding protein (MBP); N-utilization substance (NusA); small ubiquitin related modifier (SUMO) fusion tag; Streptavidin binding peptide (STREP); Tandem affinity purification (TAP); and thioredoxin (TrxA). Other tags may be used in the present invention. These and other fusion tags have been described, e.g. Costa et al. Frontiers in Microbiology 5 (2014): 63 and in PCT / US16 / 57044, the contents of which are incorporated herein by reference in their entireties. In certain embodiments, a His tag is located at SP6’s N-terminus. SP6 Promoter

[0144] Any promoter that can be recognized by an SP6 RNA polymerase may be used in the present invention. Typically, an SP6 promoter comprises 5' ATTTAGGTGACACTATAG-3' (SEQ ID NO: 10). Variants of the SP6 promoter have been discovered and / or created to optimize recognition and / or binding of SP6 to its promoter. Non-limiting variants include but are not limited to : 5'-ATTTAGGGGACACTATAGAAGAG-3": 5'- ATTTAGGGGACACTATAGAAGG-3'; 5'-ATTTAGGGGACACTATAGAAGGG-3'; 5'- ATTTAGGTGACACTATAGAA-3; 5'"-ATTTAGGTGACACTATAGAAGA-3'"; 5'- ATTTAGGTGACACTATAGAAGAG-3'; 5-ATTTAGGTGACACTATAGAAGG-3'; 5'- ATTTAGGTGACACTATAGAAGGG-3';, 5-ATTTAGGTGACACTATAGAAGNG-3"; and 5'- CATACGATTTAGGTGACACTATAG-3' (SEQ ID NO: 11 to SEQ ID NO: 20).

[0145] In addition, a suitable SP6 promoter for the present invention may be about 95%, 90%, 85%, 80%, 75%, or 70% identical or homologous to any one of SEQ ID NO: 10 to SEQ ID NO: 20. Moreover, an SP6 promoter useful in the present invention may include one or more additional nucleotides 5’ and / or 3’ to any of the promoter sequences described herein. DNA Template

[0146] Typically, a CFTR DNA template is either entirely double-stranded or mostly single-stranded with a double-stranded SP6 promoter sequence.

[0147] Linearized plasmid DNA (linearized via one or more restriction enzymes), linearized genomic DNA fragments (via restriction enzyme and / or physical means), PCR products, and / or synthetic DNA oligonucleotides can be used as templates for in vitro transcription with SP6, provided that they contain a double-stranded SP6 promoter upstream (and in the correct orientation) of the DNA sequence to be transcribed.

[0148] In some embodiments, the linearized DNA template has a blunt-end.

[0149] In some embodiments, the DNA sequence to be transcribed may be optimized to facilitate more efficient transcription and / or translation. For example, the DNA sequence may be optimized regarding cis-regulatory elements (e.g., TATA box, termination signals, and protein binding sites), artificial recombination sites, chi sites, CpG dinucleotide content, negative CpG islands, GC content, polymerase slippage sites, and / or other elements relevant to transcription; the DNA sequence may be optimized regarding cryptic splice sites, mRNA secondary structure, stable free energy of mRNA, repetitive sequences, RNA instability motif, and / or other elements relevant to mRNA processing and stability; the DNA sequence may be optimized regarding codon usage bias, codon adaptability, internal chi sites, ribosomal binding sites (e.g., IRES), premature polyA sites, Shine-Dalgarno (SD) sequences, and / or other elements relevant to translation; and / or the DNA sequence may be optimized regarding codon context, codon- anticodon interaction, translational pause sites, and / or other elements relevant to protein folding. Optimization methods known in the art may be used in the present invention, e.g., GeneOptimizer by ThermoFisher and OptimumGene™, which are described in US 20110081708, the contents of which are incorporated herein by reference in its entirety

[0150] In some embodiments, the DNA template includes a 5’ and / or 3' untranslated region. In some embodiments, a 5 untranslated region includes one or more elements that affect an mRNAs stability or translation, for example, an iron responsive element. In some embodiments, a 5’ untranslated region may be between about 50 and 500 nucleotides in length.

[0151] In some embodiments, a 3' untranslated region includes one or more of a polyadenylation signal, a binding site for proteins that affect an mRNA’s stability of location in a cell, or one or more binding sites for miRNAs. In some embodiments, a 3' untranslated region may be between 50 and 500 nucleotides in length or longer.

[0152] Exemplary 3' and / or 5’ UTR sequences can be derived from mRNA molecules which are stable (e.g., globin, actin, GAPDH, tubulin, histone, or citric acid cycle enzymes) to increase the stability of the sense mRNA molecule. For example, a 5' UTR sequence may include a partial sequence of a CMV immediate-early | (IE1) gene, or a fragment thereof to improve the nuclease resistance and / or improve the half-life of the polynucleotide. Also contemplated is the inclusion of a sequence encoding human growth hormone (hGH), or a fragment thereof to the 3' end or untranslated region of the polynucleotide (e.g.. mRNA) to further stabilize the polynucleotide. Generally, these modifications improve the stability and / or pharmacokinetic properties (e.g., half-life) of the polynucleotide relative to their unmodified counterparts, and include, for example modifications made to improve such polynucleotides’ resistance to in vivo nuclease digestion. Large-scale mRNA Synthesis

[0153] The present invention relates to large-scale production of codon optimized CFTR mRNA. In some embodiments, a method according to the invention synthesizes mRNA at least 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900mg, 1 g, Sg, 10g,25g,50g, 75g, 100g, 250 g, 500 g, 750 g, 1 kg, 5 kg, 10 kg, 50 kg, 100 kg, 1000 kg, or more at a single batch. As used herein, the term “batch” refers to a quantity or amount of mRNA synthesized at one time, e.g, produced according to a single manufacturing setting. A batch may refer to an amount of mRNA synthesized in one reaction that occurs via a single aliquot of enzyme and / or a single aliquot of DNA template for continuous synthesis under one set of conditions. mRNA synthesized at a single batch would not include mRNA synthesized at different times that are combined to achieve the desired amount. Generally, a reaction mixture includes SP6 RNA polymerase, a linear DNA template, and an RNA polymerase reaction buffer (which may include ribonucleotides or may require addition of ribonucleotides).

[0154] According to the present invention, 1-100 mg of SP6 polymerase is typically used per gram (g) of mRNA produced. In some embodiments, about 1-90 mg, 1-80 mg, 1-60 mg, 1- 50 mg, 1-40 mg, 10-100 mg, 10-80 mg, 10-60 mg, 10-50 mg of SP6 polymerase is used per gram of mRNA produced. In some embodiments, about 5-20 mg of SP6 polymerase is used to produce about 1 gram of mRNA. In some embodiments, about 0.5 to 2 grams of SP6 polymerase is used to produce about 100 grams of mRNA. In some embodiments, about 5 to 20 grams of SP6 polymerase is used to about 1 kilogram of mRNA. In some embodiments, at least 5 mg of SP6 polymerase is used to produce at least 1 gram of mRNA. In some embodiments, at least 500 mg of SP6 polymerase is used to produce at least 100 grams of mRNA. In some embodiments, at least 5 grams of SP6 polymerase is used to produce at least 1 kilogram of mRNA. In some embodiments, about 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of plasmid DNA is used per gram of mRNA produced. In some embodiments, about 10-30 mg of plasmid DNA is used to produce about 1 gram of mRNA. In some embodiments, about 1 to 3 grams of plasmid DNA is used to produce about 100 grams of mRNA. In some embodiments, about 10 to 30 grams of plasmid DNA is used to about 1 kilogram of mRNA. In some embodiments, at least 10 mg of plasmid DNA is used to produce at least 1 gram of mRNA. In some embodiments, at least 1 gram of plasmid DNA is used to produce at least 100 grams of mRNA. In some embodiments, at least 10 grams of plasmid DNA is used to produce at least | kilogram of mRNA.

[0155] In some embodiments, the concentration of the SP6 RNA polymerase in the reaction mixture may be from about 1 to 100 nM, 1 to 90 nM, 1 to 80 nM, 1 to 70 nM, 1 to 60 nM, 1 to 50 nM, 1 to 40 nM, | to 30 nM, 1 to 20 nM, or about 1 to 10 nM. In certain embodiments, the concentration of the SP6 RNA polymerase is from about 10 to 50 nM, 20 to 50 nM, or 30 to 50 nM. A concentration of 100 to 10000 Units / ml of the SP6 RNA polymerase may be used, as examples, concentrations of 100 to 9000 Units / ml, 100 to 8000 Units / ml, 100 to 7000 Units / ml. 100 to 6000 Units / ml, 100 to 5000 Units / ml, 100 to 1000 Units / ml, 200 to 2000 Units / ml, 500 to 1000 Units / ml, 500 to 2000 Units / ml, 500 to 3000 Units / ml, 500 to 4000 Units / ml, 500 to 5000 Units / ml, 500 to 6000 Units / ml, 1000 to 7500 Units / ml, and 2500 to 5000 Units / ml may be used.

[0156] The concentration of each ribonucleotide (e.g, ATP, UTP, GTP, and CTP) ina reaction mixture is between about 0.1 mM and about 10 mM, e.g, between about 1 mM and about 10 mM, between about 2 mM and about 10 mM, between about 3 mM and about 10 mM, between about 1 mM and about 8 mM, between about 1 mM and about 6 mM, between about 3 mM and about 10 mM, between about 3 mM and about 8 mM, between about 3 mM and about 6 mM, between about 4 mM and about 5 mM. In some embodiments, each ribonucleotide is at about 5S mM in a reaction mixture. In some embodiments, the total concentration of NTPs (for example, ATP, GTP, CTP and UTPs combined) used in the reaction range between 1 mM and 40 mM. In some embodiments, the total concentration of tNTPs (for example, ATP, GTP, CTP and UTPs combined) used in the reaction range between 1 mM and 30 mM, or between 1 mM and 28 mM, or between 1 mM to 25 mM, or between 1 mM and 20 mM. In some embodiments, the total tNTPs concentration is less than 30 mM. In some embodiments, the total rINTPs concentration is less than 25 mM. In some embodiments, the total rNTPs concentration is less than 20 mM. In some embodiments, the total rNTPs concentration is less than 15 mM. In some embodiments, the total INTPs concentration is less than 10 mM.

[0157] The RNA polymerase reaction buffer typically includes a salt / buffering agent, e.g., Tris, HEPES, ammonium sulfate, sodium bicarbonate, sodium citrate, sodium acetate, potassium phosphate sodium phosphate, sodium chloride, and magnesium chloride.

[0158] The pH of the reaction mixture may be between about 6 to 8.5, from 6.5 to 8.0, from 7.0 to 7.5, and in some embodiments, the pH is 7.5.

[0159] Linear or linearized DNA template (e.g, as described above and inan amount / concentration sufficient to provide a desired amount of RNA), the RNA polymerase reaction buffer, and SP6 RNA polymerase are combined to form the reaction mixture. The reaction mixture is incubated at between about 37 °C and about 42 °C for thirty minutes to six hours, e.g., about sixty to about ninety minutes.

[0160] In some embodiments, about 5 mM NTPs, about 0.05 mg / mL SP6 polymerase, and about 0.1 mg / ml DNA template in a suitable RNA polymerase reaction buffer (final reaction mixture pH of about 7.5) is incubated at about 37 °C to about 42 °C for sixty to ninety minutes.

[0161] In some embodiments, a reaction mixture contains linearized double stranded DNA template with an SP6 polymerase-specific promoter, SP6 RNA polymerase, RNase inhibitor, pyrophosphatase, 29 mM NTPs, 10 mM DTT and a reaction buffer (when at 10x is 800 mM HEPES, 20 mM spermidine, 250 mM MgCl, pH 7.7) and quantity sufficient (QS) toa desired reaction volume with RNase-free water; this reaction mixture is then incubated at 37 °C for 60 minutes. The polymerase reaction is then quenched by addition of DNase I and a DNase I buffer (when at 10x is 100 mM Tris-HCI, 5 mM MgCl: and 25 mM CaClz, pH 7.6) to facilitate digestion of the double-stranded DNA template in preparation for purification. This embodiment has been shown to be sufficient to produce 100 grams of mRNA.

[0162] Tn some embodiments, a reaction mixture includes NTPs at a concentration ranging from 1 - 10 mM, DNA template at a concentration ranging from 0.01 — 0.5 mg / ml, and SP6 RNA polymerase at a concentration ranging from 0.01 - 0.1 mg / ml, e.g, the reaction mixture comprises NTPs at a concentration of 5 mM, the DNA template at a concentration of 0.1 mg / ml, and the SP6 RNA polymerase at a concentration of 0.05 mg / ml. Nucleotides

[0163] Various naturally-occurring or modified nucleosides may be used to product mRNA according to the present invention. In some embodiments, an mRNA is or comprises natural nucleosides (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g, 2- aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5- methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5- bromouridine, CS-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methyleytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8- oxoguanosine, O(6)-methylguanine, pseudouridine, (e.g., N-1-methyl-pseudouridine), 2- thiouridine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2’-fluororibose, ribose, 2’- deoxyribose, arabinose, and hexose); and / or modified phosphate groups (e.g., phosphorothiocates and 5'-N-phosphoramidite linkages). 0164) In some embodiments, the mRNA comprises one or more nonstandard nucleotide residues. The nonstandard nucleotide residues may include, e.g, 5-methyl-cytidine (“5mC”), pseudouridine (“yU”), and / or 2-thio-uridine (“2sU”). See, e.g, U.S. Patent No. 8,278,036 or W02011012316 for a discussion of such residues and their incorporation into mRNA. The mRNA may be RNA, which is defined as RNA in which 25% of U residues are 2-thio-uridine and 25% of C residues are 5-methylcytidine. Teachings for the use of RNA are disclosed US Patent Publication US20120195936 and international publication W02011012316, both of which are hereby incorporated by reference in their entirety. The presence of nonstandard nucleotide residues may render an mRNA more stable and / or less immunogenic than a control mRNA with the same sequence but containing only standard residues. In further embodiments, the mRNA may comprise one or more nonstandard nucleotide residues chosen from isocytosine, pseudoisocytosine, S-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine and 2-chloro-6-aminopurine cytosine, as well as combinations of these modifications and other nucleobase modifications. Some embodiments may further include additional modifications to the furanose ring or nucleobase. Additional modifications may include, for example, sugar modifications or substitutions (e.g, one or more of a 2'-O-alkyl modification, a locked nucleic acid (LNA)). In some embodiments, the RNAs may be complexed or hybridized with additional polynucleotides and / or peptide polynucleotides (PNA). In some embodiments where the sugar modification is a 2'-O-alkyl modification, such modification may include, but are not limited to a 2'-deoxy-2'-fluoro modification, a 2'-O-methyl modification, a 2'-O-methoxyethyl modification and a 2'-deoxy modification. In some embodiments, any of these modifications may be present in 0-100% of the nucleotides—for example, more than 0%, 1%, 10%, 25%, 50%, 75%, 85%, 90%, 95%, or 100% of the constituent nucleotides individually or in combination. Post-synthesis processing

[0165] Typically, a 5’ cap and / or a 3' tail may be added after the synthesis. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The presence of a “tail” serves to protect the mRNA from exonuclease degradation.

[0166] A 5’ cap is typically added as follows: first, an RNA terminal phosphatase removes one of the terminal phosphate groups from the 5° nucleotide, leaving two terminal phosphates; guanosine triphosphate (GTP) is then added to the terminal phosphates via a guanylyl transferase, producing a 5°55 triphosphate linkage; and the 7-nitrogen of guanine is then methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5")ppp (5'(A,G(5")ppp(5)A and G(5")ppp(5)G. Additional cap structures are described in published US Application No. US 2016 / 0032356 and U.S. Provisional Application 62 / 464,327, filed February 27, 2017, which are incorporated herein by reference.

[0167] Typically, a tail structure includes a poly(A) and / or poly(C) tail. A poly-A or poly-C tail on the 3' terminus of mRNA typically includes at least 50 adenosine or cytosine nucleotides, at least 150 adenosine or cytosine nucleotides, at least 200 adenosine or cytosine nucleotides, at least 250 adenosine or cytosine nucleotides, at least 300 adenosine or cytosine nucleotides, at least 350 adenosine or cytosine nucleotides, at least 400 adenosine or cytosine nucleotides, at least 450 adenosine or cytosine nucleotides, at least 500 adenosine or cytosine nucleotides, at least 550 adenosine or cytosine nucleotides, at least 600 adenosine or cytosine nucleotides, at least 650 adenosine or cytosine nucleotides, at least 700 adenosine or cytosine nucleotides, at least 750 adenosine or cytosine nucleotides, at least 800 adenosine or cytosine nucleotides, at least 850 adenosine or cytosine nucleotides, at least 900 adenosine or cytosine nucleotides, at least 950 adenosine or cytosine nucleotides, or at least 1 kb adenosine or cytosine nucleotides, respectively. In some embodiments, a poly A or poly C tail may be about 10 to 800 adenosine or cytosine nucleotides (e.g., about 10 to 200 adenosine or cytosine nucleotides, about 10 to 300 adenosine or cytosine nucleotides, about 10 to 400 adenosine or cytosine nucleotides, about 10 to 500 adenosine or cytosine nucleotides, about 10 to 550 adenosine or cytosine nucleotides, about 10 to 600 adenosine or cytosine nucleotides, about 50 to 600 adenosine or cytosine nucleotides, about 100 to 600 adenosine or cytosine nucleotides, about 150 to 600 adenosine or cytosine nucleotides, about 200 to 600 adenosine or cytosine nucleotides, about 250 to 600 adenosine or cytosine nucleotides, about 300 to 600 adenosine or cytosine nucleotides, about 350 to 600 adenosine or cytosine nucleotides, about 400 to 600 adenosine or cytosine nucleotides, about 450 to 600 adenosine or cytosine nucleotides, about 500 to 600 adenosine or cytosine nucleotides, about 10 to 150 adenosine or cytosine nucleotides, about 10 to 100 adenosine or cytosine nucleotides, about 20 to 70 adenosine or cytosine nucleotides, or about 20 to 60 adenosine or cytosine nucleotides) respectively. In some embodiments, a tail structure includes is a combination of poly (A) and poly (C) tails with various lengths described herein. In some embodiments, a tail structure includes at least 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% adenosine nucleotides. In some embodiments, a tail structure includes at least 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% cytosine nucleotides.

[0168] As described herein, the addition of the 5° cap and / or the 3’ tail facilitates the detection of abortive transcripts generated during in vitro synthesis because without capping and / or tailing, the size of those prematurely aborted mRNA transcripts can be too small to be detected. Thus, in some embodiments, the 5° cap and / or the 3’ tail are added to the synthesized mRNA before the mRNA is tested for purity (e.g, the level of abortive transcripts present in the mRNA). In some embodiments, the 5° cap and / or the 3’ tail are added to the synthesized mRNA before the mRNA is purified as described herein. In other embodiments, the 5° cap and / or the 3’ tail are added to the synthesized mRNA after the mRNA is purified as described herein.

[0169] mRNA synthesized according to the present invention may be used without further purification. In particular, mRNA synthesized according to the present invention may be used without a step of removing shortmers. In some embodiments, mRNA synthesized according to the present invention may be further purified. Various methods may be used to purify mRNA synthesized according to the present invention. For example, purification of mRNA can be performed using centrifugation, filtration and / or chromatographic methods. In some embodiments, the synthesized mRNA is purified by ethanol precipitation or filtration or chromatography, or gel purification or any other suitable means. In some embodiments, the mRNA is purified by HPLC. In some embodiments, the mRNA is extracted in a standard phenol: chloroform : isoamyl alcohol solution, well known to one of skill in the art. In some embodiments, the mRNA is purified using Tangential Flow Filtration. Suitable purification methods include those described in US 2016 / 0040154, US 2015 / 0376220, PCT application PCT / US18 / 19954 entitled “METHODS FOR PURIFICATION OF MESSENGER RNA” filed on February 27, 2018, and PCT application PCT / US18 / 19978 entitled “METHODS FOR PURIFICATION OF MESSENGER RNA” filed on February 27, 2018, all of which are incorporated by reference herein and may be used to practice the present invention.

[0170] In some embodiments, the mRNA is purified before capping and tailing. In some embodiments, the mRNA is purified after capping and tailing. In some embodiments, the mRNA is purified both before and after capping and tailing.

[0171] In some embodiments, the mRNA is purified either before or after or both before and after capping and tailing, by centrifugation.

[0172] In some embodiments, the mRNA is purified either before or after or both before and after capping and tailing, by filtration.

[0173] In some embodiments, the mRNA is purified either before or after or both before and after capping and tailing, by Tangential Flow Filtration (TFF).

[0174] In some embodiments, the mRNA is purified either before or after or both before and after capping and tailing by chromatography. Characterization of mRNA

[0175] Full-length or abortive transcripts of mRNA may be detected and quantified using any methods available in the art. In some embodiments, the synthesized mRNA molecules are detected using blotting, capillary electrophoresis, chromatography, fluorescence, gel electrophoresis, HPLC, silver stain, spectroscopy, ultraviolet (UV), or UPLC, or a combination thereof. Other detection methods known in the art are included in the present invention. In some embodiments, the synthesized mRNA molecules are detected using UV absorption spectroscopy with separation by capillary electrophoresis. In some embodiments, mRNA is first denatured by a Glyoxal dye before gel electrophoresis (“Glyoxal gel electrophoresis”). In some embodiments, synthesized mRNA is characterized before capping or tailing. In some embodiments, synthesized mRNA is characterized after capping and tailing,

[0176] In some embodiments, mRNA generated by the method disclosed herein comprises less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1% impurities other than full length mRNA. The impurities include IVT contaminants, e.g., proteins, enzymes, free nucleotides and / or shortmers.

[0177] In some embodiments, mRNA produced according to the invention is substantially free of shortmers or abortive transcripts. In particular, mRNA produced according to the invention contains undetectable level of shortmers or abortive transcripts by capillary electrophoresis or Glyoxal gel electrophoresis. As used herein, the term “shortmers” or “abortive transcripts” refers to any transcripts that are less than full-length. In some embodiments, “shortmers” or “abortive transcripts” are less than 100 nucleotides in length, less than 90, less than 80, less than 70, less than 60, less than 50, less than 40, less than 30, less than 20, or less than 10 nucleotides in length. In some embodiments, shortmers are detected or quantified after adding a 5’-cap, and / or a 3 -poly A tail. mRNA Solution

[0178] In some embodiments, mRNA may be provided in a solution to be mixed with a lipid solution such that the mRNA may be encapsulated in lipid nanoparticles. A suitable mRNA solution may be any aqueous solution containing mRNA to be encapsulated at various concentrations. For example, a suitable mRNA solution may contain an mRNA ata concentration of or greater than about 0.01 mg / ml, 0.05 mg / ml, 0.06 mg / ml, 0.07 mg / ml, 0.08 mg / ml, 0.09 mg / ml, 0.1 mg / ml, 0.15 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / m}, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, or 1.0 mg / ml. In some embodiments, a suitable mRNA solution may contain an mRNA at a concentration ranging from about 0.01-1.0 mg / ml, 0.01-0.9 mg / ml, 0.01-0.8 mg / ml, 0.01-0.7 mg / ml, 0.01-0.6 mg / ml, 0.01-0.5 mg / ml, 0.01-0.4 mg / ml, 0.01-0.3 mg / ml, 0.01-0.2 mg / ml, 0.01-0.1 mg / ml, 0.05-1.0 mg / ml, 0.05-0.9 mg / ml, 0.05- 0.8 mg / ml, 0.05-0.7 mg / ml, 0.05-0.6 mg / ml, 0.05-0.5 mg / ml, 0.05-0.4 mg / ml, 0.05-0.3 mg / ml, 0.05-0.2 mg / ml, 0.05-0.1 mg / ml, 0.1-1.0 mg / ml, 0.2-0.9 mg / ml, 0.3-0.8 mg / ml, 0.4-0.7 mg / ml, or 0.5-0.6 mg / ml. In some embodiments, a suitable mRNA solution may contain an mRNA ata concentration up to about 5.0 mg / ml, 4.0 mg / ml, 3.0 mg / ml, 2.0 mg / ml, 1.0 mg / ml, .09 mg / ml, 0.08 mg / ml, 0.07 mg / ml, 0.06 mg / ml, or 0.05 mg / ml.

[0179] Typically, a suitable mRNA solution may also contain a buffering agent and / or salt. Generally, buffering agents can include HEPES, ammonium sulfate, sodium bicarbonate, sodium citrate, sodium acetate, potassium phosphate and sodium phosphate. In some embodiments, suitable concentration of the buffering agent may range from about 0.1 mM to 100 mM, 0.5 mM to 90 mM, 1.0 mM to 80 mM, 2 mM to 70 mM, 3 mM to 60 mM, 4 mM to 50 mM, 5 mM to 40 mM, 6 mM to 30 mM, 7 mM to 20 mM, 8 mM to 15 mM, or 9 to 12 mM. In some embodiments, suitable concentration of the buffering agent is or greater than about 0.1 mM, 0.5 mM, 1 mM, 2 mM, 4 mM, 6 mM, 8 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, or 50 mM.

[0180] Exemplary salts can include sodium chloride, magnesium chloride, and potassium chloride. In some embodiments, suitable concentration of salts in an mRNA solution may range from about 1 mM to S00 mM, 5 mM to 400 mM, 10 mM to 350 mM, 15 mM to 300 mM, 20 mM to 250 mM, 30 mM to 200 mM, 40 mM to 190 mM, 50 mM to 180 mM, SO mM to 170 mM, 50 mM to 160 mM, 50 mM to 150 mM, or 50 mM to 100 mM. Salt concentration in a suitable mRNA solution is or greater than about 1 mM, 5 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, or 100 mM.

[0181] In some embodiments, a suitable mRNA solution may have a pH ranging from about 3.5-6.5, 3.5-6.0, 3.5-5.5., 3.5-5.0, 3.5-4.5, 4.0-5.5, 4.0-5.0, 4.0-4.9, 4.0-4.8, 4.0-4.7, 4.0- 4.6, or 4,0-4.5. In some embodiments, a suitable mRNA solution may have a pH of or no greater than about 3.5, 4.0, 4.1,4.2,4.3,4.4,4.5,46,47,48,49,50,52,54,56,58,60,6.1,6.3, and 6.5.

[0182] Various methods may be used to prepare an mRNA solution suitable for the present invention. In some embodiments, mRNA may be directly dissolved in a buffer solution described herein. In some embodiments, an mRNA solution may be generated by mixing an mRNA stock solution with a buffer solution prior to mixing with a lipid solution for encapsulation. In some embodiments, an mRNA solution may be generated by mixing an mRNA stock solution with a buffer solution immediately before mixing with a lipid solution for encapsulation. In some embodiments, a suitable mRNA stock solution may contain mRNA in water at a concentration at or greater than about 0.2 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.8 mg / ml, 1.0 mg / ml, 1.2 mg / ml, 1.4 mg / ml, 1.5 mg / ml, or 1.6 mg / ml, 2.0 mg / ml, 2.5 mg / ml, 3.0 mg / ml, 3.5 mg / ml, 4.0 mg / ml, 4.5 mg / ml, or 5.0 mg / ml.

[0183] In some embodiments, an mRNA stock solution is mixed with a buffer solution using a pump. Exemplary pumps include but are not limited to gear pumps, peristaltic pumps and centrifugal pumps.

[0184] Typically, the buffer solution is mixed at a rate greater than that of the mRNA stock solution. For example, the buffer solution may be mixed at a rate at least 1x, 2x, 3x, 4x, 5x, 6%, 7x, 8x, 9x, 10x, 15x, or 20x greater than the rate of the mRNA stock solution. In some embodiments, a buffer solution is mixed at a flow rate ranging between about 100-6000 ml / minute (e.g., about 100-300 ml / minute, 300-600 ml / minute, 600-1200 ml / minute, 1200-2400 ml / minute, 2400-3600 ml / minute, 3600-4800 ml / minute, 4800-6000 ml / minute, or 60-420 ml / minute). In some embodiments, a buffer solution is mixed at a flow rate of or greater than about 60 ml / minute, 100 ml / minute, 140 ml / minute, 180 ml / minute, 220 ml / minute, 260 ml / minute, 300 ml / minute, 340 ml / minute, 380 ml / minute, 420 ml / minute, 480 ml / minute, 540 ml / minute, 600 ml / minute, 1200 ml / minute, 2400 ml / minute, 3600 ml / minute, 4800 ml / minute, or 6000 ml / minute.

[0185] Tn some embodiments, an mRNA stock solution is mixed at a flow rate ranging between about 10-600 ml / minute (e.g., about 5-50 ml / minute, about 10-30 ml / minute, about 30- 60 ml / minute, about 60-120 ml / minute, about 120-240 ml / minute, about 240-360 ml / minute, about 360-480 ml / minute, or about 480-600 ml / minute). In some embodiments, an mRNA stock solution is mixed at a flow rate of or greater than about 5 ml / minute, 10 ml / minute, 15 ml / minute, 20 ml / minute, 25 ml / minute, 30 ml / minute, 35 ml / minute, 40 ml / minute, 45 ml / minute, 50 ml / minute, 60 ml / minute, 80 ml / minute, 100 ml / minute, 200 ml / minute, 300 ml / minute, 400 ml / minute, 500 ml / minute, or 600 ml / minute. Delivery Vehicles

[0186] According to the present invention, mRNA encoding a CFTR protein (e.g, a full length, fragment, or portion of a CFTR protein) as described herein may be delivered as naked RNA (unpackaged) or via delivery vehicles. As used herein, the terms “delivery vehicle,” “transfer vehicle,” “nanoparticle” or grammatical equivalent, are used interchangeably. 0187) Delivery vehicles can be formulated in combination with one or more additional nucleic acids, carriers, targeting ligands or stabilizing reagents, or in pharmacological compositions where it is mixed with suitable excipients. Techniques for formulation and administration of drugs may be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa., latest edition. A particular delivery vehicle is selected based upon its ability to facilitate the transfection of a nucleic acid to a target cell.

[0188] In some embodiments, a delivery vehicle comprising CFTR mRNA is administered by pulmonary delivery, e.g., comprising nebulization. In these embodiments, the delivery vehicle may be in an aerosolized composition which can be inhaled. In some embodiments, the mRNA is expressed in the tissue in which the delivery vehicle was administered, e.g., nasal cavity, trachea, bronchi, bronchioles, and / or other pulmonary system- related cell or tissue. Additional teaching of pulmonary delivery and nebulization are described in the related international application PCT / US17 / 61100 filed November 10, 2017 by Applicant entitled “NOVEL ICE-BASED LIPID NANOPARTICLE FORMULATION FOR DELIVERY OF MRNA”, and the U. S. Provisional Application USSN 62 / 507,061, each of which is incorporated by reference in its entirety.

[0189] Tn some embodiments, mRNAs encoding a CFTR protein may be delivered via a single delivery vehicle. In some embodiments, mRNAs encoding a CFTR protein may be delivered via one or more delivery vehicles each of a different composition. According to various embodiments, suitable delivery vehicles include, but are not limited to polymer based carriers, such as polyethyleneimine (PEI), lipid nanoparticles and liposomes, nanoliposomes, ceramide-containing nanoliposomes, proteoliposomes, both natural and synthetically-derived exosomes, natural, synthetic and semi-synthetic lamellar bodies, nanoparticulates, calcium phosphor-silicate nanoparticulates, calcium phosphate nanoparticulates, silicon dioxide nanoparticulates, nanocrystalline particulates, semiconductor nanoparticulates, poly(D-arginine), sol-gels, nanodendrimers, starch-based delivery systems, micelles, emulsions, niosomes, multi- domain-block polymers (vinyl polymers, polypropyl acrylic acid polymers, dynamic polyconjugates), dry powder formulations, plasmids, viruses, calcium phosphate nucleotides, aptamers, peptides and other vectorial tags. Also contemplated is the use of bionanocapsules and other viral capsid proteins assemblies as a suitable transfer vehicle. (Hum. Gene Ther. 2008 September; 19(9):887-95).

[0190] A delivery vehicle comprising CFTR mRNA may be administered and dosed in accordance with current medical practice, taking into account the clinical condition of the subject, the site and method of administration (e.g., local and systemic, including oral, pulmonary, and via injection), the scheduling of administration, the subject's age, sex, body weight, and other factors relevant to clinicians of ordinary skill in the art. The “effective amount” for the purposes herein may be determined by such relevant considerations as are known to those of ordinary skill in experimental clinical research, pharmacological, clinical and medical arts. In some embodiments, the amount administered is effective to achieve at least some stabilization, improvement or elimination of symptoms and other indicators as are selected as appropriate measures of disease progress, regression or improvement by those of skill in the art. For example, a suitable amount and dosing regimen is one that causes at least transient protein production,

[0191] Tn some embodiments, a CFTR mRNA is administered in combination with one or more CFTR potentiators and / or correctors. Suitable CFTR potentiators and / or correctors include ivacaftor (trade name Kalydeco®), lumacaftor (trade name Orkambi®) or the combination of ivacaftor and lumacaftor. In some embodiments, a CFTR mRNA is administered in combination with one or more other CF treatment such as hormone replacement therapies, thyroid hormone replacement therapy, non-steroidal inflammatory drugs, and prescription dronabinol (Marinol®) during treatment.

[0192] In some embodiments, the human subject receives concomitant CFTR modulator therapy. In some embodiments, the concomitant CFTR modulator therapy comprises ivacaftor. In some embodiments, the concomitant CFTR modulator therapy comprises lumacaftor. In some embodiments, the concomitant CFTR modulator therapy comprises tezacaftor. In some embodiments, the concomitant CFTR modulator therapy is selected from ivacaftor, lumacaftor, tezacaftor, or a combination. In some embodiments, the concomitant CFTR modulator therapy comprises VX-659. In some embodiments, the concomitant CFTR modulator therapy comprises VX-445. In some embodiments, the concomitant CFTR modulator therapy comprises VX-152. In some embodiments, the concomitant CFTR modulator therapy comprises VX-440. In some embodiments, the concomitant CFTR modulator therapy comprises VX-371. In some embodiments, the concomitant CFTR modulator therapy comprises VX-561. In some embodiments, the concomitant CFTR modulator therapy comprises GLPG1837. In some embodiments, the concomitant CFTR modulator therapy comprises GLPG2222. In some embodiments, the concomitant CFTR modulator therapy comprises GLPG2737. In some embodiments, the concomitant CFTR modulator therapy comprises GLPG2451. In some embodiments, the concomitant CFTR modulator therapy comprises GLPG1837. In some embodiments, the concomitant CFTR modulator therapy comprises PTI-428. In some embodiments, the concomitant CFTR modulator therapy comprises PTI-801. In some embodiments, the concomitant CFTR modulator therapy comprises PTI-808. In some embodiments, the concomitant CFTR modulator therapy comprises eluforsen.

[0193] In some embodiments, the human subject is not eligible for treatment with one or more of ivacaftor, lumacaftor, tezacaftor, VX-659, VX-445, VX-152, VX-440, VX-371, VX- 561, VX-659 or combinations thereof. In some embodiments, the human subject is not eligible for treatment with one or more of ivacaftor, lumacaftor, tezacaftor, VX-659, VX-445, VX-152, VX-440, VX-371, VX-561, VX-659, GLPG1837, GLPG2222, GLPG2737, GLPG2451, GLPG1837, PTI-428, PT1-801, PTI-808, eluforsen, or combinations thereof.

[0194] In some embodiments, delivery vehicles are formulated such that they are suitable for extended-release of the mRNA contained therein. Such extended-release compositions may be conveniently administered to a subject at extended dosing intervals. Liposomal delivery vehicles

[0195] In some embodiments, a suitable delivery vehicle is a liposomal delivery vehicle, e.g, a lipid nanoparticle. As used herein, liposomal delivery vehicles, e.g., lipid nanoparticles, are usually characterized as microscopic vesicles having an interior aqua space sequestered from an outer medium by a membrane of one or more bilayers. Bilayer membranes of liposomes are typically formed by amphiphilic molecules, such as lipids of synthetic or natural origin that comprise spatially separated hydrophilic and hydrophobic domains (Lasic, Trends Biotechnol, 16: 307-321, 1998). Bilayer membranes of the liposomes can also be formed by amphiphilic polymers and surfactants (e.g., polymerosomes, niosomes, etc.). In the context of the present invention, a liposomal delivery vehicle typically serves to transport a desired mRNA to a target cell or tissue. In some embodiments, a nanoparticle delivery vehicle is a liposome. In some embodiments, a liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids. In some embodiments, a liposome comprises no more than three distinct lipid components. In some embodiments, one distinct lipid component is a sterol-based cationic lipid Cationic Lipids

[0196] As used herein, the phrase “cationic lipids” refers to any of a number of lipid species that have a net positive charge at a selected pH, such as physiological pH.

[0197] Suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2010 / 144740, which is incorporated herein by reference. In certain embodiments, the compositions and methods of the present invention include a cationic lipid, (6Z,9Z,28Z,31Z)-heptatriaconta- 6,9,28.31-tetraen-19-yl 4-(dimethylamino) butanoate, having a compound structure of: NS tS TT A ) © a Sr” TEN NN and pharmaceutically acceptable salts thereof.

[0198] Other suitable cationic lipids for use in the compositions and methods of the present invention include ionizable cationic lipids as described in International Patent Publication WO 2013 / 149140, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid of one of the following formulas: Ry i i for i re” " IN ho Ly 343 Ry \ Ly N. t re” NH » ne No or a pharmaceutically acceptable salt thereof, wherein Ry and Ra are each independently selected from the group consisting of hydrogen, an optionally substituted, variably saturated or unsaturated C1-C2o alkyl and an optionally substituted, variably saturated or unsaturated Cs-C20 acyl, wherein Li and La are each independently selected from the group consisting of hydrogen, an optionally substituted Ci-Cao alkyl, an optionally substituted variably unsaturated Ci-Cao alkenyl, and an optionally substituted C1-Cso alkynyl; wherein m and o are each independently selected from the group consisting of zero and any positive integer (e.g., where m is three); and wherein n is zero or any positive integer (e.g., where n is one). In certain embodiments, the compositions and methods of the present invention include the cationic lipid (15Z, 18Z)-N.N-dimethyl-6-(9Z,12Z)-octadeca- 9,12-dien-1-yl) tetracosa-15,18-dien-1-amine (“HGTS5000™), having a compound structure of: ~~~ | (HGT-5000) and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include the cationic lipid (15Z, 18Z)-N,N-dimethyl-6-((9Z,12Z)- octadeca-9,12-dien-1-yl) tetracosa-4,15,18-trien-l -amine (“HGTS5001”), having a compound structure of: e000 0000S (HGT-5001) and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include the cationic lipid and (15Z,18Z)-N,N-dimethyl-6- ((9Z,12Z)-octadeca-9,12-dien-1-yl) tetracosa-5,15,18-trien- 1 -amine (“HGTS002”), having a compound structure of: SAAT 3 (HGT-5002) and pharmaceutically acceptable salts thereof.

[0199] Other suitable cationic lipids for use in the compositions and methods of the invention include cationic lipids described as aminoalcohol lipidoids in International Patent Publication WO 2010 / 053572, which is incorporated herein by reference. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: Ho bo OH OH > KA CioMa¢ Crary OH Nor CioHay and pharmaceutically acceptable salts thereof.

[0200] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2016 / 118725, which is incorporated herein by reference. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of* NINN SN NNN NN | i NNN NNN NNSA i i NINN NN IN and pharmaceutically acceptable salts thereof.

[0201] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2016 / 118724, which is incorporated herein by reference. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of* ANN NTN NNN NNN 7 NTN ONT NT NT | SAN | j | on NNN NNSA NSS and pharmaceutically acceptable salts thereof.

[0202] Other suitable cationic lipids for use in the compositions and methods of the invention include a cationic lipid having the formula of 14,25-ditridecyl 15,18,21,24-tetraaza- octatriacontane, and pharmaceutically acceptable salts thereof.

[0203] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publications WO 2013 / 063468 and WO 2016 / 205691, each of which are incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: AVIA TY AIS AMR AAA H Rr: Rb AA HO NH HN ori N wy RL OH or pharmaceutically acceptable salts thereof, wherein each instance of Rl is independently optionally substituted Cs-Cao alkenyl. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: iy Cota y So) Ho INANN 10H ° SN or ey Ciotlzy Hi and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: “a = HO #4) oe — l £ : Q AnH Bs om MAA Q J T rr oF 2 and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of’ J rr (5 oH 5 a a :) 1 Q ZN; Asn HO #6 oH HAA O OH Je | 7 and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions an methods of the present invention include a cationic lipid having a compound structure of’ oF A & HO 4s ~~ Pr (ffs OH F A ZF and pharmaceutically acceptable salts thereof.

[0204] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2015 / 184256, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: HyC{CHy)m OH HyC-{CH ny Ny oN (ERR), #5 (CRaRg) 1 AR OR WL ge (CH), ~CH2 x ye HO” (CHa CHa or a pharmaceutically acceptable salt thereof, wherein each X independently is O or S; each Y independently is O or S; each m independently is 0 to 20; each n independently is 1 to 6; each Ra is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen: and each Rs is independently hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclyl, optionally substituted 3-14 membered heterocyclyl, optionally substituted C6-14 aryl, optionally substituted 5-14 membered heteroaryl or halogen. In certain embodiments, the compositions and methods of the present invention include a cationic lipid, “Target 23”, having a compound structure of: OH oO Yn Ct HCH HO», Cok Cuahar” “OH a AN HCY Q YN HO. Cf | Aon NN o el NE OH (Target 23) and pharmaceutically acceptable salts thereof.

[0205] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2016 / 004202, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: ® oo A R Foros o R® oto 0 Rag? rt Oy “ HI nN Pr 0 Rap o R= YA omAn or a pharmaceutically acceptable salt thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; o ANN TIENTS NN so NNT TIN, Q y Am w Pg og gs NO or a pharmaceutically acceptable salt thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: « ISN N= NNN 5 6 — NN } Pg St Ss Sn Se 0 or a pharmaceutically acceptable salt thereof.

[0206] Other suitable cationic lipids for use in the compositions and methods of the present invention include cationic lipids as described in United States Provisional Patent Application Serial Number 62 / 758,179, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: oO R3 x! LR Rr? = Ax WY oA mp IN . x! N A—LL LN YY R'” mT R3 O mo A R3 0 Rr? Ra x! or a pharmaceutically acceptable salt thereof, wherein each R! and R? is independently H or Ci-Cs aliphatic; each m is independently an integer having a value of 1 to 4; each A is independently a covalent bond or arylene; each L! is independently an ester, thioester, disulfide, or anhydride group; each L? is independently C2-Cho aliphatic; each X' is independently H or OH; and each R? is independently Cs-Cao aliphatic. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: C1oHz ww o YY YY AH ° Ho” H | Nom 0 CioHz1 (Compound 1) or a pharmaceutically acceptable salt thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: Bin 1 0 o Aen, ig NA SY — 4 Pa A “ny og wh A YIN | i [ST & AN HO" "CaHyr (Compound 2) or a pharmaceutically acceptable salt thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: Baa o OH I I Poy, SOY 0 wh ~SY NN a ~~ 0 NO J > (Compound 3) or a pharmaceutically acceptable salt thereof.

[0207] Other suitable cationic lipids for use in the compositions and methods of the present invention include the cationic lipids as described in J. McClellan, M. C. King, Cell 2010, 141, 210-217 and in Whitehead et al., Nature Communications (2014) 5:4277, which is incorporated herein by reference. In certain embodiments, the cationic lipids of the compositions and methods of the present invention include a cationic lipid having a compound structure of: Gratz GraHar [eXNgple] Ox y v CigHar OC atzr ~Y NTN Ay O Tr and pharmaceutically acceptable salts thereof.

[0208] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2015 / 199952, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NTN ! A Ss ts ont o ANN Mop tt ot? and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Q y | A AAAS ASS Sa NTN ~ ~ “y a eA NNN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NN SANS 1 x BNE NI NP | Pl b St tS gs tt 1 and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; NN lL ANN I NLS Qa AN NN NSN IANS he O and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NTN NONANSS I eg ANN : NPN Se Ia) and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: | NOS ANS NNN INNS ff) and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; SONS INOS ANN I : I i De o and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NNN ASSASINS NTN 8 NN OANA Y and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NNN ON ONAN } ANNAN NNN SAAN 0 and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NONTSSNN NTN ASS SSS AP pS StS © a NS and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; I~ NTN A NAN SSS J and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NTN | N, 0 ANN Se ~N ve we A, SN J and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Q L Ame AS Ny a eo Ae Ng and pharmaceutically acceptable salts thereof. 10209] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2017 / 004143, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: ONS ' AAAI NN Sa NIN INTENTS and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Id oN | AANA HENNA NN RN NN aN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Sh TNT NN TT A SN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NN ANA SN, SN Pa and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Oy SN 0 INNS i NN NN NN NNN NNN NS Pa and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: : ONS 0 AANA ena I J. NN NN NN Po No and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; fe o Angas Ap x ow © 5 MeN, SSN PONS SN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; Fa hi dail ad Oo NNN ssp ano NNN, SN PN Sse and pharmaceutically acceptable salts thereof. Tn some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: ~ ONS ° ARAN AAA gman Se, SN PN SN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: 0 A CTT NNN og SNS Mao AS a and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: BEANS CN ae “a NS ANN SSA GAA Soe, NS and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; ~ fo Fn ~ 0 NN EN Se and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: pA ALE VALIS MAW WMI RAS SL AMA AL - Aaa OS | i 0 2 ANAS o SSN 31 NNN SN Memeo SSN and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure; aN ON AAAI NS QC A ~~ hl NOAA QO and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: ANN Ba AANA OANA NNO ses NOAA O and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Ox On OS NN AANA OANA ~N ME NOS and pharmaceutically acceptable salts thereof. Tn some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NNN OT OT N oY UNTO gry fT WN Se NOAA Qo and pharmaceutically acceptable salts thereof.

[0210] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2017 / 075531, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid of the following formula: RE G3? i i ot N F; ~~ \ NN 7 2, R’ G3 Q R2 or a pharmaceutically acceptable salt thereof, wherein one of L! or L? is -O(C=0)-, -(C=0)0-, - C(=0)-, -0-, -S(O)s. -S-S-, -C(=0)S-, -SC(=0)-. -NR*C(=0)-, -C(=O)NR*-, NR*C(=0)NR*, - OC(=0)NR¥-, or -NR*C(=0)0-; and the other of L! or L? is -O(C=0)-, -(C=0)0-, -C(=0)-, -O-, - S(O) x, -S$-S-, -C(=0)S-, SC(=0)-, -NR*C(=0)-, -C(=0)NR*, NR*C(=0)NR*, -OC(=0)NR* or -NR*C(=0)0- or a direct bond; G' and G* are each independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene; G* is C1-Cas alkylene, C1-Ca4 alkenylene, Cs-Ck cycloalkylene, Ca-Cg cycloalkenylene; R* is H or C1-Ci2 alkyl; R! and R? are each independently Ce-Ca4 alkyl or Ce-Cas alkenyl; R® is H, ORS, CN, -C(=0)OR?, -OC(=0)R* or -NR® C(=0)R*; R* is C1-C12 alkyl; R® is H or C1-Ce alkyl; and x is 0, 1 or 2

[0211] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2017 / 117528, which is incorporated herein by reference. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Q SAAN GIA AAA a ~ 0 SSN Sono A Nomi A NSN and pharmaceutically acceptable salts thereof. Tn some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: NNN SSN OSS AS oA A am Ae ie O ry PN OA Q and pharmaceutically acceptable salts thereof. In some embodiments, the compositions and methods of the present invention include a cationic lipid having the compound structure: Ang Oma oO N o ASN Somos A i ml MNS and pharmaceutically acceptable salts thereof.

[0212] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2017 / 0492485, which is incorporated herein by reference. In some embodiments, the cationic lipids of the compositions and methods of the present invention include a compound of one of the following formulas: 0 PRONPN SNP PNP RNS NNN aN Fe nr RNIN NNN J NN NN 0 PY SPN NPN RNAS NNN Pa NSN ,and 0 PNP SPP PPP rN, NNN No and pharmaceutically acceptable salts thereof. For any one of these four formulas, Rs is independently selected from -(CH2)«Q and -(CH2) nCHQR; Q is selected from the group consisting of -OR, -OH, -O(CH2)uN(R)2, -OC(O)R, -CXs, -CN, -N(R)C(O)R, -N(H)C(O)R, -N(R)S(O):R, - NH)S(O):R, -N(R)C(O)N(R)z, -N(H)C(O)N(R)z, -N(H)C(O)N(H)R), -NR)C(SNR), - NH)C(S)N(R)2, -NH)C(S)N(H)(R), and a heterocycle; and n is 1, 2, or 3. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: 0 Q AeA | HOS NASA, ASA Pa and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: oO OQ PR NN NN NNN 1 LN NN NNN Pa and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: 0 AAs AN NSA * TLC and pharmaceutically acceptable salts thereof. In certain embodiments, the compositions and methods of the present invention include a cationic lipid having a compound structure of: oO AAs Ho Nass NNN Pa SS and pharmaceutically acceptable salts thereof.

[0213] Other suitable cationic lipids for use in the compositions and methods of the invention include the cationic lipids as described in International Patent Publication WO 2017 / 173054 and WO 2015 / 095340, each of which is incorporated herein by reference. Tn certain embodiments, the compositions and methods of the present invention include a cationic lipid having ...

Claims

CLAIMS We claim: 1 A pharmaceutical composition for treating cystic fibrosis, comprising an mRNA encoding a Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), wherein the CFTR comprises one or more mutations that produce an activated CFTR protein.

2. A pharmaceutical composition for treating cystic fibrosis, comprising an mRNA encoding a Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), wherein the CFTR comprises one or more mutations that produce a CFTR protein that is more stable than a wild type CFTR protein. 3 The pharmaceutical composition of claim 1 or 2, wherein the one or more mutations are located within the CFTR regulatory domain, a membrane-spanning domain (MSD), and / or a nucleotide-binding domain.

4. The pharmaceutical composition of claim 3, wherein the one or more mutations are located within the CFTR regulatory domain amine acid residues 634-835. 8, The pharmaceutical composition of any one of the preceding claims, wherein the CFTR comprises at least 2 mutations in amino acid residues in comparison to wild type CFTR protein.

6. The pharmaceutical composition of claim 5, wherein the CFTR comprises between 2 and 16 mutations in amino acid residues in comparison to wild type CFTR protein 7. The pharmaceutical composition of any one of the preceding claims, wherein the mutation is an amino acid substitution.

8. The pharmaceutical composition of claim 1 or 2, wherein the one or more mutations are made at residues K 14, K68, S422, S660, S670, S686, T690, S700, K710, S712, K716, S737, $753, 8768, T787, T788, S790, $795, $813, K978, K1041, K1080, K1218, E1371, and combinations thereof 9 The pharmaceutical composition of claim 8, wherein the one or more mutations are selected from K14R, K68R, K710R, K716R, K1041R, K1080R, K1218R, and combinations thereof.

10. The pharmaceutical composition of claim 9, wherein the CFTR comprises mutations selected from K14R, K68R, K1218R, K14R / K68R, K14R / K1218R, K68R / K1218R, and K14R / K68R / K1218R. 11 The pharmaceutical composition of claim 8, wherein the CFTR comprises mutations S660D / S737D / S795D / S813D, S660DD / S686D / S700D / S737D / S795D / S813D, S660D / S686D / S700D / S712D / S737D / S768D / S795D / S813D, K978C, or E13710Q. Bn. The pharmaceutical composition of claim 8, wherein the CFTR comprises mutations $422D / S660D / S670D / S686D / T690D / S700D / S712D / S753D / T787D / T788D / S790D / S795D / S813 D 13. The pharmaceutical composition of claim 8, wherein the CFTR comprises mutations $422D / S660D / S670D / S686D / T690D / S700D / S712D / S737D / S753D / S768D / T787D / T788D / S790 D / S795D / S813D.

14. The pharmaceutical composition of claim 8, wherein the one or more mutations made at residues $422, $660, S670, S686, T690, S700, $712, S737, $753, S768, T787, T788, S790, §795, $813 is an amino acid substitution to glutamic acid (E).

15. The pharmaceutical composition of claim 14, wherein the CFTR comprises mutations S422E / S660E / S670E / S6R6E / T690E / STO0E / S712E / S753E / T787E / T788E / S790E / S795E / S813E.

16. The pharmaceutical composition of claim 14, wherein the CFTR comprises mutations S422E / S660E / S670E / S686E / T690E / S700E / S712E / S737E / S753E / S768E / T787E / T788E / S790E / S795F / S813E.

17. The pharmaceutical composition of any one of claims 11-16, wherein the CFTR further comprises K14R mutation and / or E1371Q mutation.

18. The pharmaceutical composition of any one of the proceeding claims, wherein the mRNA encoding CFTR is codon optimized.

19. The pharmaceutical composition of claim 18, wherein the codon optimized CFTR mRNA further comprises a 5° untranslated region (UTR) sequence of SEQ ID NO: 4 20. The pharmaceutical composition of claim 18, wherein the codon optimized CFTR mRNA further comprises a 3° untranslated region (UTR) sequence of SEQ ID NO: 5 or SEQ ID NO:

6.

21. The pharmaceutical composition of any one of the preceding claims, wherein the codon optimized CFTR mRNA is encapsulated within a nanoparticle.

22. The pharmaceutical composition of claim 21, wherein the nanoparticle is a liposome.

23. The pharmaceutical composition of claim 22, wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

24. The pharmaceutical composition of claim 23, wherein the liposome comprises no more than three distinct lipid components.

25. The pharmaceutical composition of claim 24, wherein one distinct lipid component is a sterol-based cationic lipid.

26. The pharmaceutical composition of any one of claims 22-25, wherein the liposome has a size of less than about 100 nm.

27. The pharmaceutical composition of any one of the preceding claims further comprising a CFTR potentiator, corrector and / or activator.

28. The pharmaceutical composition of claim 27, wherein the pharmaceutical composition comprises a CFTR potentiator.

29. The pharmaceutical composition of claim 28, wherein the CFTR potentiator is ivacaftor.

30. A polynucleotide comprising a sequence complementary to the sequence of the mRNA of any one of the preceding claims.

31. The polynucleotide of claim 30, wherein the polynucleotide is a linear or circular polynucleotide comprising deoxyribonucleotide residues.

32. A cultured cell comprising the polynucleotide of claim 30 or 31 33. A method of inducing CFTR expression in epithelial cells in a lung of a mammal, the method comprising contacting the epithelial cells in the lung of the mammal with a composition of any one of claims 1-29.

34. The method of claim 33, wherein the codon optimized CFTR mRNA is administered via pulmonary delivery.

35. The method of claim 34, wherein the pulmonary delivery is nebulization.

36. A method of treating cystic fibrosis comprising administering to a subject in need of treatment a pharmaceutical composition of any one of claims 1-29.

37. The method of claim 36, wherein the codon optimized CFTR mRNA is administered to the subject via pulmonary delivery.

38. The method of claim 37, wherein the pulmonary delivery is nebulization.

39. The method of any one of claims 36-38, wherein treating the subject is achieved ata lower therapeutically effective dose in comparison to treating the subject with an mRNA encoding a wild type CFTR.

40. The method of claim 39, wherein treating the subject in need results in shorter nebulization times to administer a therapeutically effective dose in comparison to treating with an mRNA encoding wild type CFTR.

41. Anucleic acid encoding a modified Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein, wherein the modified CFTR protein comprises one or more amino acid substitutions to glutamic acid. 42, The nucleic acid of claim 41, wherein the one or more amino acid substitutions occur at serine residues. 43, The nucleic acid of claim 41, the one or more amino acid substitutions occur at threonine residues.

44. The nucleic acid of any one of claims 41-43, wherein the one or more amino acid substitutions are located within the CFTR regulatory domain. 45, The nucleic acid of any one of claims 41-44, wherein the one or more amino acid substitutions are located at amino acid residues 440-820.

46. The nucleic acid of claim 45, wherein the one or more amino acid substitutions occur at residue $422, S660, S670, S686, T690, $700, S712, S737, $753, $768, T787, T788, $790, S795, S813, and combinations thereof.

47. The nucleic acid of claim 41, wherein the modified CFTR protein comprises one or more mutations of S422E, S660E, S670E, S686E, T690E, S700E, S712E, S737E, S753E, S768E, T787E, T788E, S790E, S795E, S813E, and combinations thereof.

48. The nucleic acid of claim 47, wherein the modified CFTR protein comprises mutations S422F / S660FE / S670F / S686E / T690E / S700FE, / S712E / S753E / T787FE, / T788F / S790F / S795FE / S&13F.

49. The nucleic acid of claim 47, wherein the modified CFTR protein comprises mutations S422E / S660E / S670E / S686E / T690E / ST00E / S712E / S737E / S753E / S768E / T787E / T788E / S790E / S795E / S813E.

50. The nucleic acid of any one of claims 41-49, wherein the modified CFTR protein further comprises K14R mutation.

51. The nucleic acid of any one of claims 41-50, wherein the modified CFTR protein further comprises E1371Q mutation.

52. A nucleic acid encoding a modified Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein, wherein the modified CFTR protein comprises K14R and E1371Q mutations.

53. The nucleic acid of any one of claims 41-52, wherein the modified CFTR protein further comprises one or more mutations in the CFTR regulatory domain, a membrane-spanning domain (MSD), and / or a nucleotide-binding domain.

54. The nucleic acid of any one of claims 41-53, wherein the nucleic acid is DNA, cDNA, RNA, mRNA or PNA.

55. The nucleic acid of claim 54, wherein the nucleic acid is mRNA.

56. The nucleic acid of claim 55, wherein the mRNA encoding the modified CFTR protein is codon optimized.

57. The nucleic acid of claim 56, wherein the codon optimized CFTR mRNA further comprises a 5° untranslated region (UTR) sequence of SEQ ID NO: 4 58. The nucleic acid of claim 56, wherein the codon optimized CFTR mRNA further comprises a 3’ untranslated region (UTR) sequence of SEQ ID NO: 5 or SEQ ID NO:

6.

59. A pharmaceutical composition comprising any one of nucleic acids of claims 41-58.

60. The pharmaceutical composition of claim 59, wherein the nucleic acid is mRNA.

61. The pharmaceutical composition of claim 60, wherein the CFTR mRNA is encapsulated within a nanoparticle.

62. The pharmaceutical composition of claim 61, wherein the nanoparticle is a liposome.

63. The pharmaceutical composition of claim 62, wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

64. The pharmaceutical composition of claim 63, wherein the liposome comprises no more than three distinct lipid components.

65. The pharmaceutical composition of claim 64, wherein one distinct lipid component is a sterol-based cationic lipid.

66. The pharmaceutical composition of any one of claims 62-65, wherein the liposome has a size of less than about 100 nm 67. An adeno-associated virus (AAV) vector comprising any one of nucleic acids of claims 41-58 68. A modified CFTR protein comprising one or more amino acid substitutions to glutamic acid.

69. The modified CFTR protein of claim 68, wherein the one or more amino acid substitutions occur at serine residues.

70. The modified CFTR protein of claim 68, wherein the one or more amino acid substitutions occur at threonine residues.

71. The modified CFTR protein of any one of claim 68-70, wherein the one or more amino acid substitutions are located within the CFTR regulatory domain.

72. The modified CFTR protein of any one of claim 68-71, wherein the one or more amino acid substitutions are located at amino acid residues 440-820.

73. The modified CFTR protein of claim 72, wherein the one or more amino acid substitutions occur at residue S422, S660, S670, S686, T690, S700, $712, S737, S753, S768, T787, T788, S790, 8795, S813, and combinations thereof.

74. The modified CFTR protein of claim 73, wherein the modified CFTR protein comprises one or more mutations of S422E, S660E, S670E, S68GE, T690E, S700E, S712E, S737E, S753E, S768E, T787E, T788E, S790E, S795E, S813E, and combinations thereof.

78. The modified CFTR protein of claim 74, wherein the modified CFTR protein comprises mutations S422E / S660E / S670E / S686E / T690E / ST00E / S712E / S753E / T787E / T788E, / S790E / S795E / SRI3F.

76. The modified CFTR protein of claim 74, wherein the modified CFTR protein comprises mutations S422E / S660E / S670E / S686E / T690E / STO0E / S712E / S737E / S753E / S768E / T787E / T788E / S790E / S795E / S813E.

73. The modified CFTR protein of any one of claim 68-76, wherein the modified CFTR protein further comprises K14R mutation.

78. The modified CFTR protein of any one of claim 68-77, wherein the modified CFTR protein further comprises E1371Q mutation.

79. A modified CFTR protein comprising K14R and E1371Q mutations.

80. The modified CFTR protein of any one of claim 68-79, wherein the modified CFTR protein further comprise one or more mutation in the CFTR regulatory domain, a membrane- spanning domain (MSD), and / or a nucleotide-binding domain.

81. A method of inducing CFTR expression in epithelial cells in a lung of a mammal, the method comprising contacting the epithelial cells in the lung of the mammal with the composition of any one of claims 59-66.

82. The method of claim 81, wherein the composition is administered via pulmonary delivery.

83. The method of claim 82, wherein the pulmonary delivery is nebulization.

84. A method of inducing CFTR expression in a subject, the method comprising administering the AAV vector of claim 67.

85. A method of treating cystic fibrosis comprising administering to a subject in need of treatment the pharmaceutical composition of any one of claims 59-66.

86. The method of claim 85, wherein the pharmaceutical composition is administered to the subject via pulmonary delivery.

87. The method of claim 86, wherein the pulmonary delivery is nebulization.

88. A method of treating cystic fibrosis comprising administering to a subject in need of treatment the AAV vector of claim 67.

89. The method of any one of claims 85-88, wherein treating the subject is achieved ata lower therapeutically effective dose in comparison to treating the subject with a nucleic acid encoding a wild type CFTR.

90. A method of treating cystic fibrosis comprising administering to a subject in need of treatment the modified CFTR protein of any one of claims 68-80.

Images