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论文题目: One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs
英文论文题目: One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs
第一作者: Zuo, EW; Cai, YJ; Li, K; Wei, Y; Wang, BA; Sun, YD; Liu, Z; Liu, JW; Hu, XD; Wei, W; Huo, XN; Shi, LY; Tang, C; Liang, D; Wang, Y; Nie, YH; Zhang, CC; Yao, X; Wang, X; Zhou, CY; Ying, WQ; Wang, QF; Chen, RC; Shen, Q; Xu, GL; Li, JS; Sun, Q; Xiong, ZQ; Yang, H
英文第一作者: Zuo, EW; Cai, YJ; Li, K; Wei, Y; Wang, BA; Sun, YD; Liu, Z; Liu, JW; Hu, XD; Wei, W; Huo, XN; Shi, LY; Tang, C; Liang, D; Wang, Y; Nie, YH; Zhang, CC; Yao, X; Wang, X; Zhou, CY; Ying, WQ; Wang, QF; Chen, RC; Shen, Q; Xu, GL; Li, JS; Sun, Q; Xiong, ZQ; Yang, H
联系作者: Sun, Q; Xiong, ZQ; Yang, H (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Institute Neurosci,State Key Lab Neurosci,Shanghai 200031, Peoples R China.
英文联系作者: Sun, Q; Xiong, ZQ; Yang, H (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Institute Neurosci,State Key Lab Neurosci,Shanghai 200031, Peoples R China.
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发表年度: 2017
卷: 27
期: 7
页码: 933-945
摘要: The CRISPR/Cas9 system is an efficient gene-editing method, but the majority of gene-edited animals showed mosaicism, with editing occurring only in a portion of cells. Here we show that single gene or multiple genes can be completely knocked out in mouse and monkey embryos by zygotic injection of Cas9 mRNA and multiple adjacent single-guide RNAs (spaced 10-200 bp apart) that target only a single key exon of each gene. Phenotypic analysis of F0 mice following targeted deletion of eight genes on the Y chromosome individually demonstrated the robustness of this approach in generating knockout mice. Importantly, this approach delivers complete gene knockout at high efficiencies (100% on Arntl and 91% on Prrt2) in monkey embryos. Finally, we could generate a complete Prrt2 knockout monkey in a single step, demonstrating the usefulness of this approach in rapidly establishing gene-edited monkey models.
英文摘要: The CRISPR/Cas9 system is an efficient gene-editing method, but the majority of gene-edited animals showed mosaicism, with editing occurring only in a portion of cells. Here we show that single gene or multiple genes can be completely knocked out in mouse and monkey embryos by zygotic injection of Cas9 mRNA and multiple adjacent single-guide RNAs (spaced 10-200 bp apart) that target only a single key exon of each gene. Phenotypic analysis of F0 mice following targeted deletion of eight genes on the Y chromosome individually demonstrated the robustness of this approach in generating knockout mice. Importantly, this approach delivers complete gene knockout at high efficiencies (100% on Arntl and 91% on Prrt2) in monkey embryos. Finally, we could generate a complete Prrt2 knockout monkey in a single step, demonstrating the usefulness of this approach in rapidly establishing gene-edited monkey models.
刊物名称: CELL RESEARCH
英文刊物名称: CELL RESEARCH
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学科: Cell Biology
英文学科: Cell Biology
影响因子: 15.606
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论文类别: Article
英文论文类别: Article
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