Global detection of DNA repair outcomes induced by CRISPR-Cas9.
Mengzhu LiuWeiwei ZhangChangchang XinJianhang YinYafang ShangChen AiJiaxin LiFei-Long MengJiazhi HuPublished in: Nucleic acids research (2021)
CRISPR-Cas9 generates double-stranded DNA breaks (DSBs) to activate cellular DNA repair pathways for genome editing. The repair of DSBs leads to small insertions or deletions (indels) and other complex byproducts, including large deletions and chromosomal translocations. Indels are well understood to disrupt target genes, while the other deleterious byproducts remain elusive. We developed a new in silico analysis pipeline for the previously described primer-extension-mediated sequencing assay to comprehensively characterize CRISPR-Cas9-induced DSB repair outcomes in human or mouse cells. We identified tremendous deleterious DSB repair byproducts of CRISPR-Cas9 editing, including large deletions, vector integrations, and chromosomal translocations. We further elucidated the important roles of microhomology, chromosomal interaction, recurrent DSBs, and DSB repair pathways in the generation of these byproducts. Our findings provide an extra dimension for genome editing safety besides off-targets. And caution should be exercised to avoid not only off-target damages but also deleterious DSB repair byproducts during genome editing.
Keyphrases
- crispr cas
- genome editing
- dna repair
- drinking water
- dna damage
- endothelial cells
- dna damage response
- copy number
- gene expression
- single cell
- cell proliferation
- induced apoptosis
- skeletal muscle
- oxidative stress
- signaling pathway
- transcription factor
- molecular docking
- dna methylation
- circulating tumor cells
- sensitive detection
- real time pcr