Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing.
Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing.
复制标题
DOI：
10.1002/9780470942390.mo150178
复制
发表时间：
2016-03-01
影响因子：
--
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Qin W;Kutny PM;Maser RS;Dion SL;Lamont JD;Zhang Y;Perry GA;Wang H
细菌和古细菌中的CRISPR-Cas9系统最近被用于包括小鼠在内的各种模式生物的基因组编辑。CRISPR-Cas9试剂可以直接递送到小鼠受精卵中，以获得携带靶向遗传修饰的突变动物。该系统的主要组分包括提供靶特异性的指导RNA、产生DNA双链断裂的Cas9核酸酶以及携带侧接与靶位点同源的序列的预期突变的供体寡核苷酸或质粒。在这里，我们描述了使用CRISPR-Cas9系统创建转基因小鼠的一般考虑因素和实验方案。
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.