Genetic Therapy Solution for Duchenne Muscular Dystrophy
Abstract
Background: Duchenne muscular dystrophy (DMD) is a severe recessive muscle wasting disease caused by mutations in the dystrophin gene. Aim: The two major goals of this grant are to (1) develop a fully AAV encoded split CRISPR prime editing approach to edit dystrophic mice, and (2) to generate a fully AAV encoded split dystrophin approach to deliver full-length dystrophin to dystrophic mice. Approach: Using an RNA-end joining technology we have developed, two- and three-way split protein coding RNAs can be assembled from individually AAV packaged expression cassettes. In this proposal, we use the RNA end joining technology to generate AAV encodable CRISPR genome editors to correct the mdx mouse mutation and to deliver a full-length Dystrophin replacement gene. Results:(1) AAV packable CRISPR prime editor constructs were generated and validated, but proved inefficient in correcting the disease causing premature stop codon in mdx mice. A latest generation AAV packable version of a CRISPR adenosine base editor was successfully developed to correct the mdx mutation. (2) AAV packable full-length dystrophin expression vectors were successfully generated. Conclusion: The goals for this funding period have been reached. In the next funding period we will proceed to the in vivo testing stage of the experiment.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2021
- Accession Number
- AD1156567
Entities
People
- Samuel L Pfaff
Organizations
- Salk Institute for Biological Studies