基因修饰小鼠构建的革命:CRISPR/Cas9技术

The revolutionary establishment of mice with specific gene modifications: CRISPR/Cas9 technology

基因修饰小鼠在研究人类疾病致病机理和治疗手段中起到了关键作用。传统的小鼠基因组编辑使用胚胎干细胞(ES细胞),虽然可以对内源基因进行精细的修改,但是复杂、繁琐并且耗时。近几年发展的人工核酸酶可以在靶位点切割DNA双链,诱发细胞内源性修复机制,发生同源重组修复或非同源末端连接,从而提高了基因组编辑的效率。从ZFN到TALEN再到CRISPR/Cas9技术,新型基因组编辑技术正以惊人的速度渗入到生命科学的各项研究工作中。将对新型基因组编辑技术进行介绍,着重阐述CRISPR/Cas9系统介导的基因编辑技术在基因修饰小鼠中的应用,比较该系统与其他方法的优越性和不足,并对该系统进行展望。

Mice models are valuable tools for studying human disease pathogenesis and therapeutics. Traditional gene targeting in mice using embryonic stem （ES） cells, although suitable for generating sophisticated genetic modifications in endogenous genes, is complex and time-consuming. In recent years, the programmable nucleases are enable to induce site-specific DNA double-strand breaks in the genome, which enhance the efficiency of homologous recombination and/or trigger error-prone non-homologous end-joining through endogenous repair mechanisms allowing high-precision genome editing. From ZFN to TALEN and to CRISPR/Cas9, the technology of genome editing is seeping into all the fields of life sciences with amazing speed. Here, we will review the technology of targeted genome editing using programmable nucleases, especially the CRISPR/Cas9 system in mice with specific gene modifications. Furthermore, the comparison of the CRISPR/Cas9 system with other genome editing technologies and the application of this system will also be discussed.