Polynucleotides encoding methylmalonyl-CoA mutase
View Patent ↗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 lipid nanoparticle comprising a messenger RNA (mRNA) and a compound having the formula:
wherein the mRNA comprises
an open reading frame (ORF) encoding the human methylmalonyl-CoA mutase (MCM) polypeptide of SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, or SEQ ID NO:213.
2. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:213.
3. The lipid nanoparticle of claim 2 , wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO:732.
4. The lipid nanoparticle of claim 2 , wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO:732.
5. The lipid nanoparticle of claim 2 , wherein the mRNA comprises a 5′ untranslated region (UTR) comprising the nucleotide sequence of SEQ ID NO:215.
6. The lipid nanoparticle 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.
7. The lipid nanoparticle of claim 2 , wherein the mRNA comprises a poly-A tail 100 residues in length.
8. The lipid nanoparticle of claim 2 , wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.
9. The lipid nanoparticle of claim 4 , wherein the mRNA comprises a 5′ UTR comprising the nucleotide sequence of SEQ ID NO:215.
10. The lipid nanoparticle of claim 4 , 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.
11. The lipid nanoparticle of claim 4 , wherein the mRNA comprises a poly-A tail 100 residues in length.
12. The lipid nanoparticle of claim 4 , wherein at least 95% of uridines in the mRNA are 5-methoxyuridines.
13. The lipid nanoparticle of claim 4 , 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.
14. The lipid nanoparticle of claim 1 , further comprising a phospholipid, a structural lipid, and a PEG lipid.
15. The lipid nanoparticle of claim 4 , further comprising a phospholipid, a structural lipid, and a PEG lipid.
16. The lipid nanoparticle of claim 13 , further comprising a phospholipid, a structural lipid, and a PEG lipid.
17. The lipid nanoparticle of claim 14 , wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.
18. The lipid nanoparticle of claim 15 , wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.
19. The lipid nanoparticle of claim 16 , wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and the structural lipid is cholesterol.
20. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:208.
21. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:209.
22. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:210.
23. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:211.
24. The lipid nanoparticle of claim 1 , wherein the ORF encodes the human MCM polypeptide of SEQ ID NO:212.