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CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell

Skeletal muscle satellite cells (SCs) are stem cells responsible for muscle development and regeneration. Although CRISPR/Cas9 has been widely used, its application in endogenous SCs remains elusive. Here, we generate mice expressing Cas9 in SCs and achieve robust editing in juvenile SCs at the post...

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Autores principales: He, Liangqiang, Ding, Yingzhe, Zhao, Yu, So, Karl K., Peng, Xianlu L., Li, Yuying, Yuan, Jie, He, Zhiming, Chen, Xiaona, Sun, Hao, Wang, Huating
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8514971/
https://www.ncbi.nlm.nih.gov/pubmed/34534448
http://dx.doi.org/10.1016/j.stemcr.2021.08.011
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author He, Liangqiang
Ding, Yingzhe
Zhao, Yu
So, Karl K.
Peng, Xianlu L.
Li, Yuying
Yuan, Jie
He, Zhiming
Chen, Xiaona
Sun, Hao
Wang, Huating
author_facet He, Liangqiang
Ding, Yingzhe
Zhao, Yu
So, Karl K.
Peng, Xianlu L.
Li, Yuying
Yuan, Jie
He, Zhiming
Chen, Xiaona
Sun, Hao
Wang, Huating
author_sort He, Liangqiang
collection PubMed
description Skeletal muscle satellite cells (SCs) are stem cells responsible for muscle development and regeneration. Although CRISPR/Cas9 has been widely used, its application in endogenous SCs remains elusive. Here, we generate mice expressing Cas9 in SCs and achieve robust editing in juvenile SCs at the postnatal stage through AAV9-mediated short guide RNA (sgRNA) delivery. Additionally, we reveal that quiescent SCs are resistant to CRISPR/Cas9-mediated editing. As a proof of concept, we demonstrate efficient editing of master transcription factor (TF) Myod1 locus using the CRISPR/Cas9/AAV9-sgRNA system in juvenile SCs. Application on two key TFs, MYC and BCL6, unveils distinct functions in SC activation and muscle regeneration. Particularly, we reveal that MYC orchestrates SC activation through regulating 3D genome architecture. Its depletion results in strengthening of the topologically associating domain boundaries thus may affect gene expression. Altogether, our study establishes a platform for editing endogenous SCs that can be harnessed to elucidate the functionality of key regulators governing SC activities.
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spelling pubmed-85149712021-10-21 CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell He, Liangqiang Ding, Yingzhe Zhao, Yu So, Karl K. Peng, Xianlu L. Li, Yuying Yuan, Jie He, Zhiming Chen, Xiaona Sun, Hao Wang, Huating Stem Cell Reports Article Skeletal muscle satellite cells (SCs) are stem cells responsible for muscle development and regeneration. Although CRISPR/Cas9 has been widely used, its application in endogenous SCs remains elusive. Here, we generate mice expressing Cas9 in SCs and achieve robust editing in juvenile SCs at the postnatal stage through AAV9-mediated short guide RNA (sgRNA) delivery. Additionally, we reveal that quiescent SCs are resistant to CRISPR/Cas9-mediated editing. As a proof of concept, we demonstrate efficient editing of master transcription factor (TF) Myod1 locus using the CRISPR/Cas9/AAV9-sgRNA system in juvenile SCs. Application on two key TFs, MYC and BCL6, unveils distinct functions in SC activation and muscle regeneration. Particularly, we reveal that MYC orchestrates SC activation through regulating 3D genome architecture. Its depletion results in strengthening of the topologically associating domain boundaries thus may affect gene expression. Altogether, our study establishes a platform for editing endogenous SCs that can be harnessed to elucidate the functionality of key regulators governing SC activities. Elsevier 2021-09-16 /pmc/articles/PMC8514971/ /pubmed/34534448 http://dx.doi.org/10.1016/j.stemcr.2021.08.011 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
He, Liangqiang
Ding, Yingzhe
Zhao, Yu
So, Karl K.
Peng, Xianlu L.
Li, Yuying
Yuan, Jie
He, Zhiming
Chen, Xiaona
Sun, Hao
Wang, Huating
CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title_full CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title_fullStr CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title_full_unstemmed CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title_short CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
title_sort crispr/cas9/aav9-mediated in vivo editing identifies myc regulation of 3d genome in skeletal muscle stem cell
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8514971/
https://www.ncbi.nlm.nih.gov/pubmed/34534448
http://dx.doi.org/10.1016/j.stemcr.2021.08.011
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