Systematic profiling of CRISPR/Cas9 outcomes in skeletal muscle stem cells reveals mechanisms of DNA repair, indicating implications for gene therapy safety.
Key Points
The research aims to explore the in vivo outcomes of CRISPR/Cas9-mediated DNA repair in skeletal muscle stem cells.
Utilized a CRISPR/Cas9/AAV9-sgRNA system to profile editing outcomes at 95 loci in skeletal muscle stem cells.
Characterized various types of DNA repair outcomes, including editing indels and large modifications.
Disrupted NHEJ and MMEJ pathways to assess their roles in regulating large on-target modifications.
Identified small editing indels and a high occurrence of large deletions and large insertions.
Large insertions included both exogenous AAV vector integrations and endogenous genomic DNA fragments.
Endogenous large insertions preferentially originate from active genomic regions influenced by chromatin architecture.