Title |
Low incidence of SNVs and indels in trio genomes of Cas9-mediated multiplex edited sheep
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Published in |
BMC Genomics, May 2018
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DOI | 10.1186/s12864-018-4712-z |
Pubmed ID | |
Authors |
Xiaolong Wang, Jing Liu, Yiyuan Niu, Yan Li, Shiwei Zhou, Chao Li, Baohua Ma, Qifang Kou, Bjoern Petersen, Tad Sonstegard, Xingxu Huang, Yu Jiang, Yulin Chen |
Abstract |
The simplicity of the CRISPR/Cas9 system has enabled its widespread applications in generating animal models, functional genomic screening and in treating genetic and infectious diseases. However, unintended mutations produced by off-target CRISPR/Cas9 nuclease activity may lead to negative consequences. Especially, a very recent study found that gene editing can introduce hundreds of unintended mutations into the genome, and have attracted wide attention. To address the off-target concerns, urgent characterization of the CRISPR/Cas9-mediated off-target mutagenesis is highly anticipated. Here we took advantage of our previously generated gene-edited sheep and performed family trio-based whole genome sequencing which is capable of discriminating variants in the edited progenies that are inherited, naturally generated, or induced by genetic modification. Three family trios were re-sequenced at a high average depth of genomic coverage (~ 25.8×). After developing a pipeline to comprehensively analyze the sequence data for de novo single nucleotide variants, indels and structural variations from the genome; we only found a single unintended event in the form of a 2.4 kb inversion induced by site-specific double-strand breaks between two sgRNA targeting sites at the MSTN locus with a low incidence. We provide the first report on the fidelity of CRISPR-based modification for sheep genomes targeted simultaneously for gene breaks at three coding sequence locations. The trio-based sequencing approach revealed almost negligible off-target modifications, providing timely evidences of the safe application of genome editing in vivo with CRISPR/Cas9. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 29 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 6 | 21% |
Student > Bachelor | 4 | 14% |
Student > Master | 3 | 10% |
Student > Ph. D. Student | 2 | 7% |
Lecturer | 1 | 3% |
Other | 2 | 7% |
Unknown | 11 | 38% |
Readers by discipline | Count | As % |
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Agricultural and Biological Sciences | 7 | 24% |
Biochemistry, Genetics and Molecular Biology | 7 | 24% |
Veterinary Science and Veterinary Medicine | 2 | 7% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 3% |
Medicine and Dentistry | 1 | 3% |
Other | 1 | 3% |
Unknown | 10 | 34% |