Ocular Gene Exon Replacement Using AAV RNA Trans-Splicing
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Solution Overview
Problem
Current treatments are inadequate for heritable ocular diseases caused by mutations in large ocular genes like ABCA4, CEP290, and MYO7A, as existing gene therapy methods struggle with packaging limitations and inefficiencies in correcting multiple mutations.
Innovation Solution
Employing pre-RNA trans-splicing molecules (RTMs) to replace defective exons in targeted ocular genes using recombinant adeno-associated virus (rAAV) vectors, enabling ex vivo and in vivo gene correction by trans-splicing, overcoming packaging constraints and correcting mutations in genes like ABCA4, CEP290, and MYO7A.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional gene therapy approaches are used to treat inherited retinal diseases, then treatment strategies can be applied, but the complexity of repairing multiple mutations in large ocular genes makes the approach ineffective
Solution Approach 1:
The patent segments the large ocular genes into smaller functional units (exons) and targets specific mutated exons for replacement rather than attempting to repair the entire gene. This segmentation allows the therapy to focus on discrete problematic regions, making the complex task of repairing multiple mutations manageable and effective.
Solution Approach 2:
The invention extracts and removes the defective exons containing mutations from the gene structure and replaces them with healthy wild-type exon sequences. This extraction approach eliminates the harmful genetic material while preserving the functional integrity of the remaining gene, effectively addressing the complexity of multiple mutations.
2Ease of manufacture
If strategies for repairing mutations in large ocular genes are implemented, then treatment can be attempted, but the challenges make the strategies inadequate
Solution Approach 1:
The patent uses copying by creating synthetic DNA sequences that replicate the healthy wild-type exon structures and inserting them into the patient's gene. This copying approach provides accurate replacements that faithfully reproduce the functional sequences, making the repair strategy both easy to manufacture and reliable in effect.
Solution Approach 2:
The invention employs intermediary vectors or delivery mechanisms that facilitate the insertion of corrected exons into the patient's ocular cells. These intermediaries simplify the manufacturing and delivery process while ensuring reliable integration of the repaired genetic material, bridging the gap between ease of manufacture and treatment adequacy.
3Measurement precision
If multiple mutations throughout large ocular genes are present, then the genetic defects can be identified, but the complexity of repairing all mutations makes treatment challenging
Solution Approach 1:
The patent applies local quality by targeting and repairing only the specific exons containing mutations while leaving the rest of the gene intact. This localized approach maintains the precision of mutation identification and correction while reducing the overall complexity by focusing efforts only on the problematic regions rather than the entire gene structure.
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
Effectively corrects genetic defects in ocular genes, potentially treating conditions such as Stargardt disease, retinitis pigmentosa, and Usher syndrome by restoring functional gene expression and reducing disease progression or severity.
Implementation Method 1
a pre-RNA trans-splicing molecule (RTM) that can replace an exon or multiple exons in a targeted mammalian ocular gene carrying a defect or mutation causing an ocular disease with an exon(s) having the naturally-occurring sequence without the defect or mutation
Implementation Method 2
recombinant adeno-associated virus (rAAV) vectors to replace defective exons in targeted ocular genes with naturally occurring sequences
Data Source
AI summary
A nucleic acid trans-splicing molecule is provided that can replace an exon in a targeted mammalian ocular gene carrying a defect or mutation causing an ocular disease with an exon having the naturally-occurring sequence without the defect or mutation. A method of treating an ocular disease, e.g., Stargardt's Disease, caused by a defect or mutation in a target gene, e.g., ABCA4 comprising: administering to the ocular cells of a subject having an ocular disease a composition comprising a recombinant AAV comprising a nucleic acid trans-splicing molecule as described above.