Scott Greg J, Gruzdev Artiom
Knockout Mouse Core, Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, USA.
Methods Mol Biol. 2019;1960:23-40. doi: 10.1007/978-1-4939-9167-9_2.
Transgenic mouse models can be subdivided into two main categories based on genomic location: (1) targeted genomic manipulation and (2) random integration into the genome. Despite the potential confounding insertional mutagenesis and host locus-dependent expression, random integration transgenics allowed for rapid in vivo assessment of gene/protein function. Since precise genomic manipulation required the time-consuming prerequisite of first generating genetically modified embryonic stem cells, the rapid nature of generating random integration transgenes remained a strong benefit outweighing various disadvantages. The advent of targetable nucleases, such as CRISPR/Cas9, has eliminated the prerequisite of first generating genetically modified embryonic stem cells for some types of targeted genomic mutations. This chapter outlines the generation of mouse models with targeted genomic manipulation using the CRISPR/Cas9 system directly into single cell mouse embryos.
（1）靶向基因组操作和（2）随机整合到基因组中。尽管存在潜在的混杂插入诱变和宿主基因座依赖性表达，但随机整合转基因允许对基因/蛋白质功能进行快速的体内评估。由于精确的基因组操作需要首先生成基因修饰的胚胎干细胞这一耗时的前提条件，因此生成随机整合转基因的快速特性仍然是一个强大的优势，超过了各种缺点。可靶向核酸酶（如CRISPR/Cas9）的出现，消除了某些类型的靶向基因组突变首先生成基因修饰胚胎干细胞的前提条件。本章概述了使用CRISPR/Cas9系统直接对单细胞小鼠胚胎进行靶向基因组操作来生成小鼠模型的方法。
