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Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos.

Published on Feb 1, 2014in Cell36.22
· DOI :10.1016/j.cell.2014.01.027
Yuyu Niu15
Estimated H-index: 15
,
Bin Shen14
Estimated H-index: 14
(National Resource Center)
+ 25 AuthorsJiahao Sha33
Estimated H-index: 33
(Nanjing Medical University)
Abstract
Summary Monkeys serve as important model species for studying human diseases and developing therapeutic strategies, yet the application of monkeys in biomedical researches has been significantly hindered by the difficulties in producing animals genetically modified at the desired target sites. Here, we first applied the CRISPR/Cas9 system, a versatile tool for editing the genes of different organisms, to target monkey genomes. By coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, we successfully achieve precise gene targeting in cynomolgus monkeys. We also show that this system enables simultaneous disruption of two target genes ( Ppar-γ and Rag1 ) in one step, and no off-target mutagenesis was detected by comprehensive analysis. Thus, coinjection of one-cell-stage embryos with Cas9 mRNA and sgRNAs is an efficient and reliable approach for gene-modified cynomolgus monkey generation.
  • References (25)
  • Citations (584)
References25
Newest
#1Yanfang Fu (CAS: Chinese Academy of Sciences)H-Index: 14
#2Jennifer A. Foden (CAS: Chinese Academy of Sciences)H-Index: 4
Last.Jeffry D. Sander (CAS: Chinese Academy of Sciences)H-Index: 4
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#2Jun Wang (CAS: Chinese Academy of Sciences)H-Index: 7
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#2Wataru Fujii (UTokyo: University of Tokyo)H-Index: 11
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#1Masahito Watanabe (International Institute of Minnesota)H-Index: 11
#2Kazuaki Nakano (Meiji University)H-Index: 12
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#1Wakako Kumita (Central Institute for Experimental Animals)H-Index: 2
#2Kenya Sato (Central Institute for Experimental Animals)H-Index: 3
Last.Yasubumi Sakakibara (Keio: Keio University)H-Index: 29
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#1Wataru Fujii (UTokyo: University of Tokyo)H-Index: 11
#2Haruka Ito (UTokyo: University of Tokyo)
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#2Zhaohui Yang (Kunming University of Science and Technology)H-Index: 1
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#1Huan Deng (HUST: Huazhong University of Science and Technology)
#2Wei Huang (HUST: Huazhong University of Science and Technology)
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