Production of genetically engineered mice with higher efficiency, lower mosaicism, and multiplexing capability using maternally expressed Cas9
Production of genetically engineered mice with higher efficiency, lower mosaicism, and multiplexing capability using maternally expressed Cas9
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DOI：
10.1038/s41598-020-57996-7
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发表时间：
2020-01-23
影响因子：
4.6
通讯作者：
Shindo, Takayuki
中科院分区：
文献类型：
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作者：
Sakurai, Takayuki;Kamiyoshi, Akiko;Shindo, Takayuki
CRISPR/Cas9系统被广泛用于产生基因编辑的动物。在这里，我们开发了一种有效的系统，用于使用来自Cas9转基因小鼠的母体Cas9产生遗传修饰的小鼠。使用该系统，与使用外源性施用的Cas9（mRNA/蛋白质）系统获得的类似参数相比，我们实现了较低的镶嵌性和较高的敲入成功率、基因编辑率和出生率。此外，我们成功地在多个位点（最多9个）同时诱导突变。我们基于母体Cas9的新型基因编辑系统可能有助于产生具有单个和多个基因修饰的小鼠。
The CRISPR/Cas9 system is widely used to generate gene-edited animals. Here, we developed an efficient system for generating genetically modified mice using maternal Cas9 from Cas9 transgenic mice. Using this system, we achieved lower mosaicism and higher rates of knock-in success, gene-editing, and birth compared to the similar parameters obtained using exogenously administered Cas9 (mRNA/protein) system. Furthermore, we successfully induced simultaneous mutations at multiple loci (a maximum of nine). Our novel gene-editing system based on maternal Cas9 could potentially facilitate the generation of mice with single and multiple gene modifications.