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Series GSE134527 Query DataSets for GSE134527
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(single vs dual)]
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 To compare the editing efficiency between the single and dual sgRNAs treatment, a middle dose (1×1011 vg/mouse) of single AAV-sgMyoD or AAV9-dual sgMyoD virus was injected into Pax7Cas9 mice intramuscularly at P10 and administration of a middle dose of AAV9 virus containing pAAV9-sgRNA backbone without any sgRNA insertion was 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
E-mail(s) 1155068845@link.cuhk.edu.hk
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 (9)
GSM3955171 MyoD_Ctrl_mid dose_deep seq_rep1
GSM3955172 MyoD_Ctrl_mid dose_deep seq_rep2
GSM3955173 MyoD_Ctrl_mid 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
Relations
BioProject PRJNA555493
SRA SRP215384

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Supplementary file Size Download File type/resource
GSE134527_RAW.tar 113.6 Mb (http)(custom) TAR (of ZIP)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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