CRISPR/Cas9/AAV9-mediated in vivo editing identifies MYC regulation of 3D genome in skeletal muscle stem cell
Year: 2021
Type: article
Abstract: 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.
Source: Stem Cell Reports
Institution: Chinese University of Hong Kong
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,800
Awards: G1603742419, G2061137899, G2345963340, Functional characterization of Atf3 in skeletal muscle stem cells and muscle regeneration, G2458090340, YY1 deficiency in satellite cell exacerbates fibrosis in dystrophic muscle: elucidation of satellite cell/macrophage/fibro-adipogenic progenitor crosstalk in Duchenne muscular dystrophy mice, Mechanistic exploration of super-enhancer associated seRNA function in skeletal muscle cells, Functional characterization of YY1 in skeletal muscle stem cells and muscle regeneration, Identification and functional characterization of super enhancer associated RNAs (seRNAs) in skeletal myogenesis using integrated genomic approach, Elucidation of post-transcriptional regulatory functions of Dhx36 in skeletal muscle stem cells and muscle regeneration, 3D genome rewiring during muscle stem cell linage development and aging, Mechanistic investigation of Linc-p27 function in skeletal muscle satellite cell and muscle regeneration, Large scale interaction analysis of RNA binding proteins/LncRNAs to identify lncRNA nuclear localization mechanisms, Genome-wide computational identification and in vivo CRISPR screen of Key Transcription Factors (TFs) and TF hotspots governing muscle satellite cell lineage progression, Functional dissection of lncRNA SAM in skeletal muscle stem cells and muscle regeneration, G8399814013
OpenAlex ID: W3199608648