Abca4 trans-splicing molecules

Abstract

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.

What is claimed: 
     
         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.