Trans-Splicing Molecules for Compact AAV Delivery of Gene Correction
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Solution Overview
Problem
Current treatments for Stargardt Disease and Leber congenital amourosis 10 (LCA 10) face limitations due to packaging size constraints of adeno-associated viral (AAV) vectors, which hinder delivery of large nucleic acid molecules needed to correct ABCA4 and CEP290 gene mutations.
Innovation Solution
Development of nucleic acid trans-splicing molecules that operatively link a binding domain, splicing domain, and coding domain to correct mutations in the ABCA4 or CEP290 genes by trans-splicing functional exons to endogenous exons, thereby replacing nonfunctional proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If AAV vectors are used for gene therapy delivery, then safety profile is improved, but packaging size constraints worsen the delivery of large nucleic acid molecules
Solution Approach 1:
The invention divides the gene therapy approach into two segments: (1) using compact AAV vectors to deliver small trans-splicing molecules, and (2) using these molecules to correct large gene mutations (ABCA4 or CEP290) through trans-splicing. This segmentation allows the vector to remain within size constraints while still achieving the therapeutic effect of correcting large genes.
Solution Approach 2:
The trans-splicing molecules act as intermediaries between the AAV vector and the target gene. The vector delivers these intermediate molecules, which then perform the actual gene correction function through trans-splicing, allowing the vector to avoid directly carrying the large gene sequence.
2Productivity
If large nucleic acid molecules are delivered to correct gene mutations, then treatment effectiveness is improved, but vector packaging capacity is worsened
Solution Approach 1:
The invention extracts the essential corrective function from the full gene sequence and encapsulates it in compact trans-splicing molecules. These molecules contain only the necessary splicing information (binding domain, splicing domain, coding domain) to correct the mutation, rather than delivering the entire large gene sequence.
Solution Approach 2:
The invention changes the parameter of information density by using trans-splicing molecules that encode correction instructions in a compact format. The molecules use specific domain structures (binding domain for target recognition, splicing domain for splice site recognition, coding domain for functional sequence) to achieve efficient information packing that fits within vector constraints.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The trans-splicing molecules effectively correct gene mutations, potentially treating or preventing diseases associated with ABCA4 or CEP290, such as Stargardt Disease and LCA 10, by restoring functional protein expression.
Implementation Method 1
a binding domain configured to bind a target ABCA4 intron
Implementation Method 2
a splicing domain configured to mediate trans-splicing; wherein the nucleic acid trans-splicing molecule is configured to trans-splice the coding domain to an endogenous ABCA4 exon
Data Source
AI summary
The present invention features nucleic acid trans-splicing molecules (e.g., pre-mRNA trans-splicing molecules (RTMs)) capable of correcting one or more mutations in the ABCA4 gene or the CEP290 gene. Such molecules are useful in the treatment of disorders associated with mutations in ABCA4, such as Stargardt Disease (e.g., Stargardt Disease 1) and disorders associated with a mutation in CEP290, such as Leber congenital amourosis 10 (LCA 10). Also provided by the invention described herein are methods of using the nucleic acid trans-splicing molecules for correcting mutations in ABCA4 and CEP290 and for treating disorders associated with mutations in ABCA4 and CEP290, such as Stargardt Disease and LCA 10.