AAV-delivered CRISPR/Cas9 for CEP290 gene editing
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Solution Overview
Problem
Current therapies for CEP290-associated diseases, such as Leber Congenital Amaurosis 10, are limited due to the difficulty in delivering gene therapy vectors for large protein sequences and the lack of approved small molecule treatments, necessitating a more effective genome editing approach.
Innovation Solution
The use of CRISPR/Cas9 genome editing systems, delivered via adeno-associated viral vectors, to target and correct the CEP290 gene mutations in retinal cells by administering nucleic acids encoding Cas9 and guide RNAs, specifically designed to alter the CEP290 gene sequence, thereby restoring functional protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If gene therapy vectors are used to deliver large protein sequences, then the therapeutic effect is improved, but the difficulty of packaging and delivering the vectors increases
Solution Approach 1:
The patent divides the CEP290 gene correction task into two separate deliverable components: a Cas9 nuclease (either as protein or encoded by a compact gene) and guide RNAs. This segmentation allows each component to be packaged into separate AAV vectors, circumventing the packaging limit problem of delivering the entire 90kb CEP290 gene sequence while achieving the same therapeutic goal of correcting the IVS26 mutation.
2Manufacturing precision
If CRISPR/Cas9 genome editing is used to correct CEP290 mutations, then the editing precision is improved, but the complexity of the delivery system increases
Solution Approach 1:
The patent uses adeno-associated viral (AAV) vectors as intermediary delivery vehicles to transport the CRISPR/Cas9 components (Cas9 and guide RNAs) into retinal cells. The AAV vectors serve as a bridge between the external administration and the intracellular CRISPR editing machinery, simplifying the overall delivery system while maintaining high editing precision through the specificity of guide RNA-targeted Cas9 nucleases.
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
This method achieves high levels of productive editing in retinal cells, with up to 50% of photoreceptor cells showing productive alterations, effectively addressing the genetic defect and potentially restoring sensory cilia function.
Implementation Method 1
the inventors have developed a strategy for correcting the IVS26 mutation in cells using CRISPR/Cas9 genome editing
Implementation Method 2
delivered via adeno-associated viral vectors, to target and correct the CEP290 gene mutations in retinal cells
Data Source
AI summary
Nucleic acids and viral vectors, particularly adeno-associated virus (AAV) vectors are provided that encode Cas9 and paired guide RNAs. The nucleic acids and vectors, and compositions that comprise them, can be used in methods to treat subjects, to alter cells in subjects who may suffer from an inherited retinal dystrophy such as CEP290 associated disease or who may be in need of alteration of a cell or a cellular nucleic acid sequence associated with an inherited retinal dystrophy such as the CEP290 gene, and/or to treat inherited retinal dystrophies including CEP290 associated disease.