Qin Wenning, Wang Haoyi
The Jackson Laboratory, Bar Harbor, ME, USA.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2019;1874:179-190. doi: 10.1007/978-1-4939-8831-0_10.
The CRISPR-Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including the mice. In this scheme, components of the CRISPR-Cas9 system are delivered into the mouse zygote and mutant mice carrying genetic modifications derived. Although microinjection has been the technology of choice, electroporation has also emerged and been proven to be effective delivering CRISPR-Cas9 reagents into the mouse zygote. Here, we describe the experimental protocol employing electroporation to deliver CRISPR-Cas9 reagents into mouse embryos and derive gene-edited mutant mice.
细菌和古细菌中的CRISPR-Cas9系统最近已被用于各种模式生物(包括小鼠)的基因组编辑。在该方案中,CRISPR-Cas9系统的组件被导入小鼠受精卵中,并由此获得携带基因修饰的突变小鼠。尽管显微注射一直是首选技术,但电穿孔法也已出现并被证明能有效地将CRISPR-Cas9试剂导入小鼠受精卵。在此,我们描述了采用电穿孔法将CRISPR-Cas9试剂导入小鼠胚胎并获得基因编辑突变小鼠的实验方案。