Truncated Dysferlin Gene for AAV Delivery
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Solution Overview
Problem
Current methods for treating dysferlinopathy, a muscular dystrophy caused by mutations in the dysferlin gene, are ineffective in slowing muscle function loss or reversing the dystrophic phenotype due to the large size of the dysferlin protein, which exceeds the packaging capacity of a single adeno-associated viral (AAV) vector.
Innovation Solution
Development of a truncated dysferlin gene that can be packaged in a single AAV vector, retaining at least a portion of the biological activity of wild-type dysferlin, allowing for effective delivery and expression of dysferlin in cells and tissues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the full-length dysferlin gene is used for AAV-mediated gene therapy, then the complete biological activity of dysferlin is achieved, but the gene size exceeds the packaging capacity of a single AAV vector
Solution Approach 1:
The invention extracts and removes specific non-essential domains (C2D and C2F) from the full-length dysferlin protein sequence, creating a truncated version that fits within AAV packaging capacity while retaining core biological functions. This extraction principle directly resolves the contradiction by reducing gene size without completely sacrificing biological activity.
Solution Approach 2:
The dysferlin protein is segmented into essential and non-essential domains, with the C2D and C2F domains being removed while retaining other functional domains. This segmentation allows the gene to be divided into a manageable size for AAV delivery while preserving the critical functional segments necessary for biological activity.
2Ease of operation
If a truncated dysferlin gene is used to fit AAV packaging capacity, then the gene can be delivered via single AAV vector, but there is concern about retaining sufficient biological activity
Solution Approach 1:
The invention applies local quality by selectively removing specific domains (C2D and C2F) while preserving other domains that are critical for biological function. This targeted approach ensures that the truncated protein maintains sufficient biological activity in the regions that matter most for dysferlin's function, while still achieving the size reduction needed for AAV delivery.
3Quantity of substance
If multiple AAV capsids are used to deliver oversized dysferlin gene, then the complete gene can be delivered, but the approach is less efficient and requires complex assembly by host enzymes
Solution Approach 1:
By extracting and removing the C2D and C2F domains, the invention creates a compact dysferlin gene version that fits within a single AAV capsid. This eliminates the need for complex multi-capasid approaches and the associated host enzyme assembly requirements, directly resolving the contradiction between complete gene delivery and system complexity.
Data Source
AI summary
This invention relates to a truncated dysferlin nucleic acid and protein, vectors (e.g., adeno-associated virus vectors) comprising the nucleic acid and methods of using the same for delivery of dysferlin to a cell or a subject and treating dysferlinopathy.