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Potential effects of endogenous RNA/DNA hybrids on CRISPR-Cas9-mediated homology-directed repair - PubMed

7 hours ago
  • #Homology-directed repair
  • #CRISPR-Cas9
  • #R-loops
  • R-loops, endogenous RNA/DNA hybrids, may influence CRISPR-Cas9-mediated homology-directed repair (HDR) efficiency.
  • In proliferating hepatocyte-derived cells, HDR efficiency is reduced at R-loop-enriched sites, but not in quiescent hepatocytes in vivo.
  • Reducing R-loop levels via RNaseH1 overexpression or G1 arrest induction increases HDR efficiency at these sites.
  • T cell activation correlates with elevated R-loop accumulation, suggesting potential impacts on ex vivo genome editing.
  • Indel frequencies remain comparable across sites with different R-loop abundance, indicating specificity to HDR effects.