Modified polynucleotides for the production of cytoplasmic and cytoskeletal proteins
The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.
1. An mRNA encoding SEQ ID NO: 4947 and wherein said mRNA comprises a coding region having at least 80% identity to SEQ ID NO: 31871.
2. The mRNA of claim 1 which is purified.
3. The mRNA of claim 1 , wherein the mRNA comprises two stop codons.
4. The mRNA isolated polynucleotide of claim 1 , wherein the mRNA comprises a first flanking region comprising a 5′ untranslated region (UTR) and a second flanking region comprising a 3′ untranslated region (UTR), said first flanking region located at the 5′ terminus of said first region and said second flanking region located at the 3′ terminus of said first region.
5. The mRNA of claim 4 , wherein the mRNA comprises a 3′ tailing sequence of linked nucleosides at the 3′ terminus of the 3′ UTR, wherein the 3′ tailing sequence of linked nucleosides is selected from the group consisting of a poly-A tail of approximately 160 nucleotides and a poly A-G quartet.
6. The mRNA of claim 4 , wherein the 5′UTR and the 3′UTR are not derived from the same species.
7. The mRNA of claim 4 , wherein at least one of the 5′UTR or the 3′UTR is not derived from beta-globin.
8. The mRNA of claim 4 , wherein at least one of said 5′UTR or said 3′UTR is not derived from the native untranslated region of said polypeptide of interest.
9. The mRNA of claim 1 , wherein the coding region is selected from the group consisting of SEQ ID NO: 31871, 31856, 31806, 31860, 31861, 31827, 31844, 31857, 31876, 31882 and 31873.
10. A pharmaceutical composition comprising the mRNA of claim 1 .
11. The pharmaceutical composition of claim 10 , wherein the pharmaceutical composition comprises a pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is selected from a solvent, aqueous solvent, non-aqueous solvent, dispersion media, diluent, dispersion, suspension aid, surface active agent, isotonic agent, thickening or emulsifying agent, preservative, lipid, lipidoids liposome, lipid nanoparticle, core-shell nanoparticles, polymer, lipoplex peptide, protein, cell, hyaluronidase, and mixtures thereof.
12. The pharmaceutical composition of claim 11 , where the pharmaceutical composition comprises a lipid and wherein said lipid is selected from DLin-DMA, DLin-K-DMA, DLin-KC2-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DODMA, DSDMA, DLenDMA, reLNPs, PLGA and PEGylated lipids and mixtures thereof.
13. A method of producing a polypeptide of interest in a mammalian cell, tissue or organism comprising administering to said cell, tissue or organism the pharmaceutical composition of claim 10 .
14. The method of claim 13 , wherein the mRNA is formulated.
15. The method of claim 14 , wherein the formulation comprises a lipid which is selected from one of DLin-DMA, DLin-K-DMA, DLin-KC2-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DODMA, DSDMA, DLenDMA, reLNPs, PLGA, PEGylated lipids and mixtures or combinations thereof.