Title | Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy. |
Publication Type | Journal Article |
Year of Publication | 2021 |
Authors | A Pickar-Oliver, V Gough, JD Bohning, S Liu, JN Robinson-Hamm, H Daniels, WH Majoros, G Devlin, A Asokan, and CA Gersbach |
Journal | Molecular Therapy : the Journal of the American Society of Gene Therapy |
Volume | 29 |
Start Page | 3243 |
Issue | 11 |
Pagination | 3243 - 3257 |
Date Published | 11/2021 |
Abstract | Targeted gene-editing strategies have emerged as promising therapeutic approaches for the permanent treatment of inherited genetic diseases. However, precise gene correction and insertion approaches using homology-directed repair are still limited by low efficiencies. Consequently, many gene-editing strategies have focused on removal or disruption, rather than repair, of genomic DNA. In contrast, homology-independent targeted integration (HITI) has been reported to effectively insert DNA sequences at targeted genomic loci. This approach could be particularly useful for restoring full-length sequences of genes affected by a spectrum of mutations that are also too large to deliver by conventional adeno-associated virus (AAV) vectors. Here, we utilize an AAV-based, HITI-mediated approach for correction of full-length dystrophin expression in a humanized mouse model of Duchenne muscular dystrophy (DMD). We co-deliver CRISPR-Cas9 and a donor DNA sequence to insert the missing human exon 52 into its corresponding position within the DMD gene and achieve full-length dystrophin correction in skeletal and cardiac muscle. Additionally, as a proof-of-concept strategy to correct genetic mutations characterized by diverse patient mutations, we deliver a superexon donor encoding the last 28 exons of the DMD gene as a therapeutic strategy to restore full-length dystrophin in >20% of the DMD patient population. This work highlights the potential of HITI-mediated gene correction for diverse DMD mutations and advances genome editing toward realizing the promise of full-length gene restoration to treat genetic disease. |
DOI | 10.1016/j.ymthe.2021.09.003 |
Short Title | Molecular Therapy : the Journal of the American Society of Gene Therapy |