Zheng Wu, Gu Feng
State Key Laboratory Cultivating Base of Optometry and Visual Science, Key Laboratory of Vision Science, Ministry of Health, The Eye Hospital of Wenzhou Medical University, Wenzhou 325027 China.
Yi Chuan. 2015 Oct;37(10):1003-10. doi: 10.16288/j.yczz.15-070.
The clustered regulatory interspaced short palindromic repeat/Cas9 (CRISPR/Cas9) system mediates genome editing and is revolutionizing genetic researches. Scientists are able to manipulate the gene of interest from any organism with CRISPR/Cas9. Compared with zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) technologies, the CRISPR/Cas9 technology provides an easy and efficient approach to manipulate the genome. In this system, sgRNA (Single guide RNA), a short RNA matching the targeted DNA fragment, guides the CRISPR/Cas9 to interrogate the genome. Because sgRNA can tolerate certain mismatches to the DNA targets and thereby promote undesired off-target mutagenesis, the key limit of this technology is off-target effects. To eliminate the off-target effects, different strategies have been adopted. In this review, we summarize the application of CRISPR/Cas9 and different strategies for addressing off-target effects.
成簇规律间隔短回文重复序列/Cas9（CRISPR/Cas9）系统介导基因组编辑，正在彻底改变基因研究。科学家能够利用CRISPR/Cas9操纵任何生物体中感兴趣的基因。与锌指核酸酶（ZFNs）和转录激活样效应物核酸酶（TALENs）技术相比，CRISPR/Cas9技术提供了一种简便高效的基因组操纵方法。在该系统中，sgRNA（单向导RNA），一种与靶向DNA片段匹配的短RNA，引导CRISPR/Cas9对基因组进行检测。由于sgRNA能够耐受与DNA靶点的某些错配，从而促进意外的脱靶诱变，该技术的关键限制是脱靶效应。为了消除脱靶效应已采用了不同策略。在本综述中，我们总结了CRISPR/Cas9的应用以及解决脱靶效应的不同策略。
