ABCA4 TRANS-SPLICING MOLECULES
Provided herein are nucleic acid trans-splicing molecules (e.g., pre-mRNA trans-splicing molecules (RTMs); RNA exon editing molecules) capable of correcting mutations in the ABCA4 gene. Such molecules are useful in the treatment of disorders such as ABCA4-associated retinal dystrophies (e.g., Stargardt Disease or cone-rod dystrophy). Also described herein are methods of using the nucleic acid trans-splicing molecules described herein to correct mutations in ABCA4, thereby treating disorders associated with mutations in ABCA4 and use of the nucleic acid trans-splicing molecules described herein for treating disorders associated with mutations in ABCA4 and in the preparation of medicaments for the treatment of disorders associated with mutations in ABCA4.
1 .- 30 . (canceled)
31 . A nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a linker domain comprising SEQ ID NO: 27 or a sequence having at least 90% identity to SEQ ID NO: 27; and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA.
32 . The nucleic acid trans-splicing molecule of claim 31 , wherein the linker domain is located between the coding domain sequence and the binding domain.
33 . The nucleic acid trans-splicing molecule of claim 31 , wherein the coding domain sequence comprises one or more ABCA4 exons and wherein the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.
34 . A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a linker domain comprising SEQ ID NO: 27 or a sequence having at least 90% identity to SEQ ID NO: 27; and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA, wherein the endogenous pre-mRNA comprises at least one mutation associated with the disease or disorder.
35 . The method of claim 34 , wherein the linker domain is located between the coding domain sequence and the binding domain.
36 . The method of claim 34 , wherein the disease or disorder is an ABCA4-associated retinal dystrophy, the coding domain sequence comprises one or more ABCA4 exons, and the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.
37 . A nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a potentiator domain comprising a U1-binding site; and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA.
38 . The nucleic acid trans-splicing molecule of claim 37 , wherein the potentiator domain comprises SEQ ID NO: 61 or a sequence having 90% to SEQ ID NO: 61.
39 . The nucleic acid trans-splicing molecule of claim 37 , wherein the potentiator domain comprises SEQ ID NO: 62 or a sequence having 90% to SEQ ID NO: 62.
40 . The nucleic acid trans-splicing molecule of claim 37 , wherein the potentiator domain is between the coding domain sequence and the binding domain.
41 . The nucleic acid trans-splicing molecule of claim 37 , wherein the coding domain sequence comprises one or more ABCA4 exons and wherein the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.
42 . A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a potentiator domain comprising a U1-binding site; and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA, wherein the endogenous pre-mRNA comprises at least one mutation associated with the disease or disorder.
43 . The method of claim 42 , wherein the potentiator domain comprises SEQ ID NO: 61 or a sequence having 90% to SEQ ID NO: 61.
44 . The method of claim 42 , wherein the potentiator domain comprises SEQ ID NO: 62 or a sequence having 90% to SEQ ID NO: 62.
45 . The method of claim 42 , wherein the potentiator domain is between the coding domain sequence and the binding domain.
46 . The method of claim 42 , wherein the disease or disorder is an ABCA4-associated retinal dystrophy, the coding domain sequence comprises one or more ABCA4 exons, and the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.
47 . A nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a translational potentiator domain comprising an AU-rich element (ARE); and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA.
48 . The nucleic acid trans-splicing molecule of claim 47 , wherein the ARE comprises SEQ ID NO: 63 or a sequence having at least 90% identity to SEQ ID NO: 63.
49 . The nucleic acid trans-splicing molecule of claim 47 , wherein the ARE is between the coding domain sequence and the binding domain.
50 . The nucleic acid trans-splicing molecule of claim 47 , wherein the coding domain sequence comprises one or more ABCA4 exons and the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.
51 . A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a nucleic acid trans-splicing molecule comprising:
(a) a coding domain sequence;
(b) a translational potentiator domain comprising an AU-rich element (ARE); and
(c) a binding domain that is complementary to a binding site within an endogenous pre-mRNA, wherein the endogenous pre-mRNA comprises at least one mutation associated with the disease or disorder.
52 . The method of claim 51 , wherein the ARE comprises SEQ ID NO: 63 or a sequence having at least 90% identity to SEQ ID NO: 63.
53 . The method of claim 51 , wherein the ARE is between the coding domain sequence and the binding domain.
54 . The method of claim 51 , wherein the disease or disorder is an ABCA4-associated retinal dystrophy, the coding domain sequence comprises one or more ABCA4 exons, and the endogenous pre-mRNA is an endogenous ABCA4 pre-mRNA.