MODIFIED POLYNUCLEOTIDES FOR TREATING PROTEIN DEFICIENCY
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 a modified mRNA, wherein said modified mRNA encodes a member selected from the group consisting of iduronidase, alpha-L-(SEQ ID NO: 1080), iduronidase, alpha-L-(SEQ ID NO: 1081), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 992), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 993), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 994), iduronate 2-sulfatase (SEQ ID NO: 1073), iduronate 2-sulfatase (SEQ ID NO: 1074), iduronate 2-sulfatase (SEQ ID NO: 1075), iduronate 2-sulfatase (SEQ ID NO: 1076), iduronate 2-sulfatase (SEQ ID NO: 1077), iduronate 2-sulfatase (SEQ ID NO: 1078), iduronate 2-sulfatase (SEQ ID NO: 1079), arylsulfatase B (SEQ ID NO: 845), arylsulfatase B (SEQ ID NO: 847) and arylsulfatase B (SEQ ID NO: 848), and wherein said modified mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.
2 . The method of claim 1 , wherein the modified mRNA comprises the modification 1-methylpseudouridine and further comprises the modification 5-methylcytosine.
3 . The method of claim 1 , wherein the modified mRNA comprises the modifications 5-methylcytosine and 2-thiouridine, wherein 25% of the cytosine residues in the modified mRNA are 5-methylcytosines and 25% of the uridine residues in the modified mRNA are 2-thiouridine.
4 . The method of claim 1 , wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the modified mRNA is not derived from beta-globin.
5 . A modified mRNA encoding a polypeptide of interest, said polypeptide of interest selected from the group consisting of iduronidase, alpha-L- (SEQ ID NO: 1080), iduronidase, alpha-L- (SEQ ID NO: 1081), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 992), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 993), galactosamine (N-acetyl)-6-sulfate sulfatase (SEQ ID NO: 994), iduronate 2-sulfatase (SEQ ID NO: 1073), iduronate 2-sulfatase (SEQ ID NO: 1074), iduronate 2-sulfatase (SEQ ID NO: 1075), iduronate 2-sulfatase (SEQ ID NO: 1076), iduronate 2-sulfatase (SEQ ID NO: 1077), iduronate 2-sulfatase (SEQ ID NO: 1078), iduronate 2-sulfatase (SEQ ID NO: 1079), arylsulfatase B (SEQ ID NO: 845), arylsulfatase B (SEQ ID NO: 847) and arylsulfatase B (SEQ ID NO: 848), and wherein said modified mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.
6 . The modified mRNA of claim 5 , wherein the modified mRNA comprises the modification 1-methylpseudouridine and further comprises the modification 5-methylcytosine.
7 . The modified mRNA of claim 5 , wherein the modified mRNA comprises the modifications 5-methylcytosine and 2-thiouridine, wherein 25% of the cytosine residues in the modified mRNA are 5-methylcytosines and 25% of the uridine residues in the modified mRNA are 2-thiouridine.
8 . The modified mRNA of claim 5 , wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the modified mRNA is not derived from beta-globin.
9 . A method of treating a disease or disorder associated with a polypeptide deficiency in a mammalian subject comprising administering to said subject the modified mRNA of claim 5 .