Polynucleotides encoding methylmalonyl-CoA mutase
The disclosure relates to polynucleotides comprising an open reading frame of linked nucleosides encoding human methylmalonyl-CoA mutase precursor, human methylmalonyl-CoA mutase (MCM) mature form, or functional fragments thereof. In some embodiments, the disclosure includes methods of treating methylmalonic acidemia in a subject in need thereof comprising administering an mRNA encoding an MCM polypeptide.
1. A messenger RNA (mRNA) comprising
an open reading frame (ORF) encoding the human methylmalonyl-CoA mutase (MCM) polypeptide of SEQ ID NO:213, wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO:732.
2. The mRNA of claim 1 , wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO:732.
3. The mRNA of claim 1 , wherein the mRNA comprises a 5′ untranslated region (UTR) comprising the nucleotide sequence of SEQ ID NO:215.
4. The mRNA of claim 1 , wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl.
5. The mRNA of claim 1 , wherein the mRNA comprises a poly-A tail 100 residues in length.
6. The mRNA of claim 1 , wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.
7. The mRNA of claim 2 , wherein the mRNA comprises a 5′ UTR comprising the nucleotide sequence of SEQ ID NO:215.
8. The mRNA of claim 2 , wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl.
9. The mRNA of claim 2 , wherein the mRNA comprises a poly-A tail 100 residues in length.
10. The mRNA of claim 2 , wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.
11. The mRNA of claim 2 , wherein the mRNA comprises a 5′ UTR comprising the nucleotide sequence of SEQ ID NO:215, wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl, wherein the mRNA comprises a poly-A tail 100 residues in length, and wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.
12. A pharmaceutical composition comprising the mRNA of claim 1 and a pharmaceutically acceptable excipient.
13. A pharmaceutical composition comprising the mRNA of claim 2 and a pharmaceutically acceptable excipient.
14. A pharmaceutical composition comprising the mRNA of claim 11 and a pharmaceutically acceptable excipient.
15. A lipid nanoparticle comprising the mRNA of claim 1 .
16. A lipid nanoparticle comprising the mRNA of claim 2 .
17. A lipid nanoparticle comprising the mRNA of claim 11 .
18. A pharmaceutical composition comprising the lipid nanoparticle of claim 15 .
19. A pharmaceutical composition comprising the lipid nanoparticle of claim 16 .
20. A pharmaceutical composition comprising the lipid nanoparticle of claim 17 .
21. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the mRNA of claim 1 .
22. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the mRNA of claim 2 .
23. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the mRNA of claim 11 .
24. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 12 .
25. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 13 .
26. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 14 .
27. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the lipid nanoparticle of claim 15 .
28. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the lipid nanoparticle of claim 16 .
29. A method of treating methylmalonic acidemia in a human subject in need thereof, the method comprising administering to the human subject an effective amount of the lipid nanoparticle of claim 17 .