Modified polynucleotides for treating protein deficiency
View Patent ↗The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.
1. A method of producing a polypeptide of interest in a mammalian cell or tissue comprising contacting said mammalian cell or tissue with an mRNA having a 3′ untranslated region and a 5′ untranslated region, wherein said mRNA encodes heparan-alpha-glucosaminide N-acetyltransferase (SEQ ID NO: 48935), and wherein said mRNA comprises a coding region, said coding region having at least 85% identity to SEQ ID NO: 84333.
2. The method of claim 1 , wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the mRNA is not derived from beta-globin.
3. The method of claim 1 , wherein the mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.
4. The method of claim 3 , wherein the mRNA comprises the modification 5-methylcytosine and 2-thiouridine.
5. The method of claim 4 , wherein 25% of the cytosine residues in the mRNA are 5-methylcytosines and 25% of the uridine residues in the mRNA are 2-thiouridine.
6. The method of claim 3 , wherein the mRNA comprises the modification pseudouridine and 5-methylcytosine.
7. The method of claim 3 , wherein the mRNA comprises the modification 1-methylpseudouridine.
8. The method of claim 7 , wherein the mRNA further comprises the modification 5-methylcytosine.
9. An mRNA encoding heparan-alpha-glucosaminide N-acetyltransferase (SEQ ID NO: 48935), said mRNA having a 3′ untranslated region and a 5′ untranslated region and wherein said mRNA comprises a coding region, said coding region having at least 85% identity to SEQ ID NO: 84333.
10. The mRNA of claim 9 , wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the mRNA is not derived from beta-globin.
11. The mRNA of claim 9 , wherein the mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.
12. The mRNA of claim 11 , wherein the mRNA comprises the modification 5-methylcytosine and 2-thiouridine.
13. The mRNA of claim 12 , wherein 25% of the cytosine residues in the mRNA are 5-methylcytosines and 25% of the uridine residues in the mRNA are 2-thiouridine.
14. The mRNA of claim 11 , wherein the mRNA comprises the modification pseudouridine and 5-methylcytosine.
15. The mRNA of claim 11 , wherein the mRNA comprises the modification 1-methylpseudouridine.
16. The mRNA of claim 15 , wherein the mRNA further comprises the modification 5-methylcytosine.