MODIFIED NUCLEIC ACIDS, AND ACUTE CARE USES THEREOF
The invention provides compositions and methods for effecting wound healing in a mammal, where the compositions include therapeutic mRNA which incorporate modified nucleosides and nucleotides.
1 . A synthetic isolated RNA comprising:
(a) a first region of linked nucleosides encoding a polypeptide of interest, said polypeptide of interest selected from the group consisting of SEQ ID NOS 86-170;
(b) a first terminal region located at the 5′ terminus of said first region comprising a 5′ untranslated region (UTR);
(c) a second terminal region located at the 3′ terminus of said first region comprising a 3′ UTR; and
(d) a 3′ tailing region of linked nucleosides;
wherein any of the regions (a)-(d) comprise at least one modified nucleoside.
2 . The synthetic isolated RNA of claim 1 wherein the at least one modified nucleoside is not 5-methylcytosine or pseudouridine.
3 . The synthetic isolated RNA of claim 1 , wherein the 5′ UTR is the native 5′UTR of the encoded polypeptide of interest.
4 . The synthetic isolated RNA of claim 1 , wherein the first terminal region comprises at least one 5′ cap structure.
5 . The synthetic isolated RNA of claim 4 , wherein the at least one 5′ cap structure is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′ fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, 2-azido-guanosine, Cap2 and Cap4.
6 . The synthetic isolated RNA of claim 1 , wherein the 5′UTR comprises a translation initiation sequence selected from the group consisting of Kozak sequence and an internal ribosome entry site (IRES).
7 . The synthetic isolated RNA of claim 1 , wherein the 3′UTR is the native 3′UTR of the encoded polypeptide of interest.
8 . The synthetic isolated RNA of claim 1 , wherein the 3′ tailing region is selected from the group consisting of a PolyA tail and PolyA-G quartet.
9 . The synthetic isolated RNA of claim 4 , wherein the 3′ tailing region is a PolyA tail and the PolyA tail is approximately 150 to 170 nucleotides in length.
10 . The synthetic isolated RNA of claim 9 , wherein the PolyA tail is approximately 160 nucleotides in length.
11 . The synthetic isolated RNA of claim 10 , which is purified.
12 . A method of treating a mammalian subject in need thereof comprising administering the synthetic isolated RNA of claim 11 .
13 . The method of claim 12 , wherein the mammalian subject is suffering from or is at risk of developing an acute or life-threatening disease or condition.
14 . The method of claim 13 , wherein the mammalian subject is suffering from a traumatic injury.
15 . The method of claim 13 , wherein the polypeptide of interest accelerates wound healing.