Alternative nucleic acid molecules and uses thereof
View Patent ↗The present disclosure provides alternative nucleosides, nucleotides, and nucleic acids, and methods of using them.
1. An mRNA encoding a polypeptide, wherein at least one base is 1-methyl-pseudouracil and one base is 5-methoxy-uracil.
2. A pharmaceutical composition comprising the mRNA of claim 1 and a pharmaceutically acceptable excipient.
3. A method of expressing a polypeptide in a mammalian cell, said method comprising the steps of:
(i) providing an mRNA of claim 1 ; and
(ii) introducing said mRNA to a mammalian cell under conditions that permit the expression of the polypeptide by the mammalian cell,
thereby expressing the polypeptide in the mammalian cell.
4. The mRNA of claim 1 , wherein about 25% of the uracils are 1-methyl-pseudouracil and about 75% of the uracils are 5-methoxy-uracil.
5. The mRNA of claim 1 , wherein about 50% of the uracils are 1-methyl-pseudouracil and about 50% of the uracils are 5- methoxy-uracil.
6. The mRNA of claim 1 , wherein about 75% of the uracils are 1-methyl-pseudouracil and about 25% of the uracils are 5-methoxy-uracil.
7. The mRNA of claim 1 , wherein at least one base is 5-methyl-cytosine.
8. The mRNA of claim 1 , wherein at least one nucleotide is alpha-thio-adenosine or alpha-thio-guanosine.
9. The mRNA of claim 8 , wherein about 2-100% of the adenosines and/or guanosines in the mRNA are alpha-thio-adenosine and/or alpha-thio-guanosine.
10. The mRNA of claim 9 , wherein about 2% of the adenosines and/or guanosines in the mRNA are alpha-thio-adenosine and/or alpha-thio-guanosine.
11. The mRNA of claim 1 further comprising:
(i) at least one 5′-cap structure;
(ii) a 5′-UTR optionally comprising a Kozak sequence; and
(iii) a 3′-UTR.
12. The mRNA of claim 11 , wherein the at least one 5′-cap structure is Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, or 2-azido-guanosine.
13. The mRNA of claim 11 further comprising a poly-A tail.
14. The mRNA of claim 1 , wherein the mRNA is codon optimized.