Skip to main content
Journal cover image

In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy.

Publication ,  Journal Article
Kwon, JB; Ettyreddy, AR; Vankara, A; Bohning, JD; Devlin, G; Hauschka, SD; Asokan, A; Gersbach, CA
Published in: Mol Ther Methods Clin Dev
December 11, 2020

Delivery of therapeutic transgenes with adeno-associated viral (AAV) vectors for treatment of myopathies has yielded encouraging results in animal models and early clinical studies. Although certain AAV serotypes efficiently target muscle fibers, transduction of the muscle stem cells, also known as satellite cells, is less studied. Here, we used a Pax7nGFP;Ai9 dual reporter mouse to quantify AAV transduction events in satellite cells. We assessed a panel of AAV serotypes for satellite cell tropism in the mdx mouse model of Duchenne muscular dystrophy and observed the highest satellite cell labeling with AAV9 following local or systemic administration. Subsequently, we used AAV9 to interrogate CRISPR/Cas9-mediated gene editing of satellite cells in the Pax7nGFP;mdx mouse. We quantified the level of gene editing using a Tn5 transposon-based method for unbiased sequencing of editing outcomes at the Dmd locus. We also found that muscle-specific promoters can drive transgene expression and gene editing in satellite cells. Lastly, to demonstrate the functionality of satellite cells edited at the Dmd locus by CRISPR in vivo, we performed a transplantation experiment and observed increased dystrophin-positive fibers in the recipient mouse. Collectively, our results confirm that satellite cells are transduced by AAV and can undergo gene editing to restore the dystrophin reading frame in the mdx mouse.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Mol Ther Methods Clin Dev

DOI

ISSN

2329-0501

Publication Date

December 11, 2020

Volume

19

Start / End Page

320 / 329

Location

United States

Related Subject Headings

  • 3206 Medical biotechnology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Kwon, J. B., Ettyreddy, A. R., Vankara, A., Bohning, J. D., Devlin, G., Hauschka, S. D., … Gersbach, C. A. (2020). In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy. Mol Ther Methods Clin Dev, 19, 320–329. https://doi.org/10.1016/j.omtm.2020.09.016
Kwon, Jennifer B., Adarsh R. Ettyreddy, Ashish Vankara, Joel D. Bohning, Garth Devlin, Stephen D. Hauschka, Aravind Asokan, and Charles A. Gersbach. “In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy.Mol Ther Methods Clin Dev 19 (December 11, 2020): 320–29. https://doi.org/10.1016/j.omtm.2020.09.016.
Kwon JB, Ettyreddy AR, Vankara A, Bohning JD, Devlin G, Hauschka SD, et al. In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy. Mol Ther Methods Clin Dev. 2020 Dec 11;19:320–9.
Kwon, Jennifer B., et al. “In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy.Mol Ther Methods Clin Dev, vol. 19, Dec. 2020, pp. 320–29. Pubmed, doi:10.1016/j.omtm.2020.09.016.
Kwon JB, Ettyreddy AR, Vankara A, Bohning JD, Devlin G, Hauschka SD, Asokan A, Gersbach CA. In Vivo Gene Editing of Muscle Stem Cells with Adeno-Associated Viral Vectors in a Mouse Model of Duchenne Muscular Dystrophy. Mol Ther Methods Clin Dev. 2020 Dec 11;19:320–329.
Journal cover image

Published In

Mol Ther Methods Clin Dev

DOI

ISSN

2329-0501

Publication Date

December 11, 2020

Volume

19

Start / End Page

320 / 329

Location

United States

Related Subject Headings

  • 3206 Medical biotechnology