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AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice.

Publication ,  Journal Article
Nance, ME; Shi, R; Hakim, CH; Wasala, NB; Yue, Y; Pan, X; Zhang, T; Robinson, CA; Duan, SX; Yao, G; Yang, NN; Chen, S-J; Wagner, KR; Duan, D ...
Published in: Molecular therapy : the journal of the American Society of Gene Therapy
September 2019

CRISPR editing of muscle stem cells (MuSCs) with adeno-associated virus serotype-9 (AAV9) holds promise for sustained gene repair therapy for muscular dystrophies. However, conflicting evidence exists on whether AAV9 transduces MuSCs. To rigorously address this question, we used a muscle graft model. The grafted muscle underwent complete necrosis before regenerating from its MuSCs. We injected AAV9.Cre into Ai14 mice. These mice express tdTomato upon Cre-mediated removal of a floxed stop codon. About 28%-47% and 24%-89% of Pax7+ MuSCs expressed tdTomato in pre-grafts and regenerated grafts (p > 0.05), respectively, suggesting AAV9 efficiently transduced MuSCs, and AAV9-edited MuSCs renewed successfully. Robust MuSC transduction was further confirmed by delivering AAV9.Cre to Pax7-ZsGreen-Ai14 mice in which Pax7+ MuSCs are genetically labeled by ZsGreen. Next, we co-injected AAV9.Cas9 and AAV9.gRNA to dystrophic mdx mice to repair the mutated dystrophin gene. CRISPR-treated and untreated muscles were grafted to immune-deficient, dystrophin-null NSG.mdx4cv mice. Grafts regenerated from CRISPR-treated muscle contained the edited genome and yielded 2.7-fold more dystrophin+ cells (p = 0.015). Importantly, increased dystrophin expression was not due to enhanced formation of revertant fibers or de novo transduction by residual CRISPR vectors in the graft. We conclude that AAV9 effectively transduces MuSCs. AAV9 CRISPR editing of MuSCs may provide enduring therapy.

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Published In

Molecular therapy : the journal of the American Society of Gene Therapy

DOI

EISSN

1525-0024

ISSN

1525-0016

Publication Date

September 2019

Volume

27

Issue

9

Start / End Page

1568 / 1585

Related Subject Headings

  • Transduction, Genetic
  • Satellite Cells, Skeletal Muscle
  • Regeneration
  • RNA, Guide, CRISPR-Cas Systems
  • Myoblasts
  • Muscular Dystrophy, Duchenne
  • Muscle, Skeletal
  • Mice, Knockout
  • Mice
  • Genetic Vectors
 

Citation

APA
Chicago
ICMJE
MLA
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Nance, M. E., Shi, R., Hakim, C. H., Wasala, N. B., Yue, Y., Pan, X., … Duan, D. (2019). AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice. Molecular Therapy : The Journal of the American Society of Gene Therapy, 27(9), 1568–1585. https://doi.org/10.1016/j.ymthe.2019.06.012
Nance, Michael E., Ruicheng Shi, Chady H. Hakim, Nalinda B. Wasala, Yongping Yue, Xiufang Pan, Tracy Zhang, et al. “AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice.Molecular Therapy : The Journal of the American Society of Gene Therapy 27, no. 9 (September 2019): 1568–85. https://doi.org/10.1016/j.ymthe.2019.06.012.
Nance ME, Shi R, Hakim CH, Wasala NB, Yue Y, Pan X, et al. AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice. Molecular therapy : the journal of the American Society of Gene Therapy. 2019 Sep;27(9):1568–85.
Nance, Michael E., et al. “AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice.Molecular Therapy : The Journal of the American Society of Gene Therapy, vol. 27, no. 9, Sept. 2019, pp. 1568–85. Epmc, doi:10.1016/j.ymthe.2019.06.012.
Nance ME, Shi R, Hakim CH, Wasala NB, Yue Y, Pan X, Zhang T, Robinson CA, Duan SX, Yao G, Yang NN, Chen S-J, Wagner KR, Gersbach CA, Duan D. AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice. Molecular therapy : the journal of the American Society of Gene Therapy. 2019 Sep;27(9):1568–1585.

Published In

Molecular therapy : the journal of the American Society of Gene Therapy

DOI

EISSN

1525-0024

ISSN

1525-0016

Publication Date

September 2019

Volume

27

Issue

9

Start / End Page

1568 / 1585

Related Subject Headings

  • Transduction, Genetic
  • Satellite Cells, Skeletal Muscle
  • Regeneration
  • RNA, Guide, CRISPR-Cas Systems
  • Myoblasts
  • Muscular Dystrophy, Duchenne
  • Muscle, Skeletal
  • Mice, Knockout
  • Mice
  • Genetic Vectors