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

Author:

Zuo Erwei12ORCID,Sun Yidi3ORCID,Wei Wu456ORCID,Yuan Tanglong2ORCID,Ying Wenqin1ORCID,Sun Hao7,Yuan Liyun4ORCID,Steinmetz Lars M.589ORCID,Li Yixue41011ORCID,Yang Hui1ORCID

Affiliation:

1. Institute 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.

2. Center for Animal Genomics, Agricultural Genome Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518124, China.

3. CAS 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.

4. Key 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.

5. Stanford Genome Technology Center, Stanford University, Palo Alto, CA 94304, USA.

6. Center for Biomedical Informatics, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai 200040, China.

7. University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0726, USA.

8. Department of Genetics, School of Medicine, Stanford University, Stanford, CA 94305, USA.

9. Genome Biology Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.

10. Collaborative Innovation Center for Genetics and Development, Fudan University, Shanghai 200438, China.

11. Shanghai Center for Bioinformation Technology, Shanghai Industrial Technology Institute, Shanghai 201203, China.

Abstract

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

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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