Simple Genome Editing of Rodent Intact Embryos by Electroporation.
Simple Genome Editing of Rodent Intact Embryos by Electroporation.
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DOI：
10.1371/journal.pone.0142755
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Mashimo T
中科院分区：
文献类型：
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作者：
Kaneko T;Mashimo T
簇状规则间隔短回文重复(CRISPR)/CRISPR相关(Cas)系统是动物基因组编辑的有力工具。最近，已经开发了新的技术来修改动物的基因，而不需要使用高技能的技术，例如原核显微注射核酸内切酶。动物电穿孔基因敲除技术(Take)是一种通过电穿孔技术将核酸内切酶mRNAs导入完整胚胎产生基因敲除大鼠的简单而有效的技术。本研究采用Take方法和CRISPR/Cas系统，成功地建立了基因敲除和基因敲入小鼠和大鼠。电穿孔Cas9mRNA、gRNA和单链寡核苷酸(SsODN)的小鼠和大鼠胚胎中编辑的靶基因包括敲除(67%的小鼠和88%的大鼠)或敲入(均为33%)。Take方法可以作为一种强大的工具，通过基因组编辑来生产转基因动物。
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE) method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN) comprised the edited targeted gene as a knockout (67% of mice and 88% of rats) or knock-in (both 33%). The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.