MODIFIED POLYNUCLEOTIDES FOR TREATING LIPASE A, LYSOSOMAL ACID, CHOLESTEROL ESTERASE 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 lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1127), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1128), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1129), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1130), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1131), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1132), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1133), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1134), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1135), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1136) and lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1137), 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 lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1127), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1128), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1129), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1130), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1131), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1132), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1133), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1134), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1135), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1136) and lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1137), 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 . (canceled)