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Deprecated: Implicit conversion from float 265.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Cell+Res 2017 ; 27 (7): 933-45 Nephropedia Template TP
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One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs #MMPMID28585534
Zuo E; Cai YJ; Li K; Wei Y; Wang BA; Sun Y; Liu Z; Liu J; Hu X; Wei W; Huo X; Shi L; Tang C; Liang D; Wang Y; Nie YH; Zhang CC; Yao X; Wang X; Zhou C; Ying W; Wang Q; Chen RC; Shen Q; Xu GL; Li J; Sun Q; Xiong ZQ; Yang H
Cell Res 2017[Jul]; 27 (7): 933-45 PMID28585534show ga
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.