Takemoto Tatsuya
Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan.
Setsuro Tech Inc., Tokushima, Japan.
Methods Mol Biol. 2020;2050:121-126. doi: 10.1007/978-1-4939-9740-4_13.
The CRISPR/Cas9 system is a powerful tool for generation of genetically modified mice. In conventional protocols, Cas9 protein (or mRNA) and sgRNA are introduced into zygotes by microinjection. However, microinjection requires special skill and is too time-consuming to treat zygotes on a large scale. Recently, we have developed a simple electroporation method which generates genetically modified mice with high efficiency. Here, we describe our method GEEP (genome editing by electroporation of Cas9 protein). This method facilitates high-throughput genetic analysis of the mouse. This chapter describes the GEEP method to generate genetically modified mice.
CRISPR/Cas9系统是用于生成基因编辑小鼠的强大工具。在传统方案中，通过显微注射将Cas9蛋白（或mRNA）和sgRNA导入受精卵。然而，显微注射需要特殊技能，且处理大量受精卵耗时过长。最近，我们开发了一种简单的电穿孔方法，可高效生成基因编辑小鼠。在此，我们描述我们的GEEP方法（通过Cas9蛋白电穿孔进行基因组编辑）。该方法有助于对小鼠进行高通量基因分析。本章描述了使用GEEP方法生成基因编辑小鼠的过程。
