Cytosine base editor generates substantial off-target single-nucleotide variants in mouse embryos

American Association for the Advancement of Science (AAAS) - Tập 364 Số 6437 - Trang 289-292 - 2019
Erwei Zuo1,2, Yidi Sun3, Wei Wu4,5,6, Tanglong Yuan1, Wenqin Ying2, Hao Sun7, Liyun Yuan5, Lars M. Steinmetz8,9,6, Yixue Li10,5,11, Hui Yang2
1Center for Animal Genomics, Agricultural Genome Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518124, China.
2Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
3CAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China.
4Center for Biomedical Informatics, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai 200040, China.
5Key Lab of Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
6Stanford Genome Technology Center, Stanford University, Palo Alto, CA 94304, USA
7University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0726, USA.
8Department of Genetics, School of Medicine, Stanford University, Stanford, CA 94305, USA
9Genome Biology Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
10Collaborative Innovation Center for Genetics and Development, Fudan University, Shanghai 200438, China.
11Shanghai Center for Bioinformation Technology, Shanghai Industrial Technology Institute, Shanghai 201203, China.

Tóm tắt

Spotting off-targets from gene editing Unintended genomic modifications limit the potential therapeutic use of gene-editing tools. Available methods to find off-targets generally do not work in vivo or detect single-nucleotide changes. Three papers in this issue report new methods for monitoring gene-editing tools in vivo (see the Perspective by Kempton and Qi). Wienert et al. followed the recruitment of a DNA repair protein to DNA breaks induced by CRISPR-Cas9, enabling unbiased detection of off-target editing in cellular and animal models. Zuo et al. identified off-targets without the interference of natural genetic heterogeneity by injecting base editors into one blastomere of a two-cell mouse embryo and leaving the other genetically identical blastomere unedited. Jin et al. performed whole-genome sequencing on individual, genome-edited rice plants to identify unintended mutations. Cytosine, but not adenine, base editors induced numerous single-nucleotide variants in both mouse and rice. Science , this issue p. 286 , p. 289 , p. 292 ; see also p. 234

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