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Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.

Publication ,  Journal Article
Pickar-Oliver, A; Gough, V; Bohning, JD; Liu, S; Robinson-Hamm, JN; Daniels, H; Majoros, WH; Devlin, G; Asokan, A; Gersbach, CA
Published in: Mol Ther
November 3, 2021

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.

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Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

November 3, 2021

Volume

29

Issue

11

Start / End Page

3243 / 3257

Location

United States

Related Subject Headings

  • Virus Integration
  • Myocardium
  • Mutation
  • Muscular Dystrophy, Duchenne
  • Muscle, Skeletal
  • Mice, Transgenic
  • Mice
  • Humans
  • Genetic Vectors
  • Genetic Therapy
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Pickar-Oliver, A., Gough, V., Bohning, J. D., Liu, S., Robinson-Hamm, J. N., Daniels, H., … Gersbach, C. A. (2021). Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy. Mol Ther, 29(11), 3243–3257. https://doi.org/10.1016/j.ymthe.2021.09.003
Pickar-Oliver, Adrian, Veronica Gough, Joel D. Bohning, Siyan Liu, Jacqueline N. Robinson-Hamm, Heather Daniels, William H. Majoros, Garth Devlin, Aravind Asokan, and Charles A. Gersbach. “Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.Mol Ther 29, no. 11 (November 3, 2021): 3243–57. https://doi.org/10.1016/j.ymthe.2021.09.003.
Pickar-Oliver A, Gough V, Bohning JD, Liu S, Robinson-Hamm JN, Daniels H, et al. Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy. Mol Ther. 2021 Nov 3;29(11):3243–57.
Pickar-Oliver, Adrian, et al. “Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy.Mol Ther, vol. 29, no. 11, Nov. 2021, pp. 3243–57. Pubmed, doi:10.1016/j.ymthe.2021.09.003.
Pickar-Oliver A, Gough V, Bohning JD, Liu S, Robinson-Hamm JN, Daniels H, Majoros WH, Devlin G, Asokan A, Gersbach CA. Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy. Mol Ther. 2021 Nov 3;29(11):3243–3257.

Published In

Mol Ther

DOI

EISSN

1525-0024

Publication Date

November 3, 2021

Volume

29

Issue

11

Start / End Page

3243 / 3257

Location

United States

Related Subject Headings

  • Virus Integration
  • Myocardium
  • Mutation
  • Muscular Dystrophy, Duchenne
  • Muscle, Skeletal
  • Mice, Transgenic
  • Mice
  • Humans
  • Genetic Vectors
  • Genetic Therapy