RT Journal Article
SR Electronic
T1 Global detection of DNA repair outcomes induced by CRISPR-Cas9
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.02.15.431335
DO 10.1101/2021.02.15.431335
A1 Mengzhu Liu
A1 Weiwei Zhang
A1 Changchang Xin
A1 Jianhang Yin
A1 Yafang Shang
A1 Chen Ai
A1 Jiaxin Li
A1 Fei-long Meng
A1 Jiazhi Hu
YR 2021
UL http://biorxiv.org/content/early/2021/02/16/2021.02.15.431335.abstract
AB 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, plasmid 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.Competing Interest StatementThe authors have declared no competing interest.