Qin Wenning, Kutny Peter M, Maser Richard S, Dion Stephanie L, Lamont Jeffrey D, Zhang Yingfan, Perry Greggory A, Wang Haoyi
The Jackson Laboratory, Bar Harbor, Maine.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Curr Protoc Mouse Biol. 2016 Mar 1;6(1):39-66. doi: 10.1002/9780470942390.mo150178.
The CRISPR-Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR-Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double-strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR-Cas9 system.
细菌和古生菌中的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。CRISPR-Cas9试剂可直接导入小鼠受精卵，以获得携带靶向基因修饰的突变动物。该系统的主要组成部分包括提供靶标特异性的引导RNA、产生DNA双链断裂的Cas9核酸酶，以及携带预期突变且两侧带有与靶位点同源序列的供体寡核苷酸或质粒。在此，我们描述了使用CRISPR-Cas9系统创建基因编辑小鼠的一般注意事项和实验方案。
