GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
Series GSE134528 Query DataSets for GSE134528
Status Public on Oct 27, 2021
Title CRISPR/Cas9/AAV9-sgRNA Mediated In Vivo Genome Editing Reveals the Indispensability of Myc During Muscle Stem Cells Activation by Remodeling the 3D Chromatin [Deep-seq(dose)]
Organism Mus musculus
Experiment type Other
Summary Skeletal muscle satellite cells (SCs) are muscle stem cells responsible for muscle development and injury induced muscle regeneration. The pace of SC related study, however, is constrained partially by the technological limitations in generating genetically modified mice. Although the ease of use of CRISPR-Cas9 in genome manipulation has been documented in many cell lines and various species, its application in endogenous SCs remains elusive. In this study, we generated muscle-specific Cas9-expressing mice and achieved robust in vivo genome editing in juvenile SCs at the postnatal stage through AAV9 mediated short guide RNAs (sgRNAs) delivery. We also found adult quiescent SCs are reluctant to CRISPR/Cas9 editing despite efficient AAV9 transduction. To edit juvenile SCs in vivo, as a proof-of-concept, we delivered sgRNAs targeting MyoD, a key gene critical for muscle physiology and showed an efficient editing at MyoD locus, resulting in accumulation of SCs and defects in SCs differentiation which resembled the phenotypes reported in MyoD knockout mice. Further application of this system on potential key transcription factors (TFs) involved in SC fate transition, Myc, Bcl6 and Pknox2, unveiled their distinct functions in the early stage of SC activation and injury induced muscle regeneration. In addition, we revealed that Myc orchestrated SCs activation through impinging on 3D chromatin architecture. Altogether we established a robust muscle restricted CRISPR/Cas9-based gene editing platform in endogenous SCs and elucidated the functionality of key factors governing SC activities.
Overall design Pax7Cas9 mice were administrated with different doses of single AAV-sgMyoD (high: 5×1011 vg/mouse; middle; 1×1011 vg/mouse; low: 0.2×1011 vg/mouse) virus intramuscularly at P10. Pax7Cas9 mice administrated with a high (5×1011 vg/mouse) dose of AAV9 virus containing pAAV9-sgRNA backbone without any sgRNA insertion were used as control. Muscle stem cells were isolated 4 weeks after injection and sgMyoD targeted locus was amplified from genomic DNA in freshly isolated satellite cells and subjected to deep sequencing.
Contributor(s) Huating W, Hao S, Liangqiang H, Yingzhe D
Citation(s) 34534448
Submission date Jul 19, 2019
Last update date Oct 28, 2021
Contact name Yingzhe Ding
Organization name The Chinese University of Hong Kong
Department Chemical Pathology
Street address Rm503,Li Ka Shing Medical Science Build.,Prince of Wales Hosp.,30-32 Ngan SHing St.
City Hong Kong
State/province Shatin NT.
ZIP/Postal code 999077
Country Hong Kong
Platforms (1)
GPL18480 Illumina HiSeq 1500 (Mus musculus)
Samples (12)
GSM3955180 MyoD_Ctrl_high dose_deep seq_rep1
GSM3955181 MyoD_Ctrl_high dose_deep seq_rep2
GSM3955182 MyoD_Ctrl_high dose_deep seq_rep3
This SubSeries is part of SuperSeries:
GSE134529 CRISPR/Cas9/AAV9-sgRNA Mediated In Vivo Genome Editing Reveals the Indispensability of Myc During Muscle Stem Cells Activation by Remodeling the 3D Chromatin
BioProject PRJNA555494
SRA SRP215385

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE134528_RAW.tar 30.7 Mb (http)(custom) TAR (of ZIP) 6.8 Mb (ftp)(http) ZIP 14.0 Mb (ftp)(http) ZIP 6.1 Mb (ftp)(http) ZIP
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap