Whole genomic analysis reveals atypical non-homologous off-target large structural variants induced by CRISPR-Cas9-mediated genome editing
Year: 2023
Type: article
Abstract: CRISPR-Cas9 genome editing has promising therapeutic potential for genetic diseases and cancers, but safety could be a concern. Here we use whole genomic analysis by 10x linked-read sequencing and optical genome mapping to interrogate the genome integrity after editing and in comparison to four parental cell lines. In addition to the previously reported large structural variants at on-target sites, we identify heretofore unexpected large chromosomal deletions (91.2 and 136 Kb) at atypical non-homologous off-target sites without sequence similarity to the sgRNA in two edited lines. The observed large structural variants induced by CRISPR-Cas9 editing in dividing cells may result in pathogenic consequences and thus limit the usefulness of the CRISPR-Cas9 editing system for disease modeling and gene therapy. In this work, our whole genomic analysis may provide a valuable strategy to ensure genome integrity after genomic editing to minimize the risk of unintended effects in research and clinical applications.
Source: Nature Communications
Authors: Hsiu-Hui Tsai, Hsiao-Jung Kao, Ming-Wei Kuo, Chin-Hsien Lin, Chun-Min Chang, Yi-Yin Chen, Hsiao-Huei Chen, Pui-Yan Kwok, Alice L. Yu, John Yu
Institutions: Chang Gung Memorial Hospital, Linkou Chang Gung Memorial Hospital, Institute of Biomedical Sciences, Academia Sinica, Academia Sinica, National Taiwan University, National Taiwan University Hospital, University of California, San Francisco, University of California San Diego, Genomics Research Center, Academia Sinica, Institute of Cellular and Organismic Biology, Academia Sinica
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,290
Award: G7084107866
OpenAlex ID: W4386167331