Wang Gan-cheng, Ming Ma, Ye Yan-zhen, Xi Jian-zhong
Department of Biomedicine, College of Engineering, Peking University, Beijing 100871, China.
1. Department of Biomedicine, College of Engineering, Peking University, Beijing 100871, China; 2. State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Molecular Medicine, Peking University, Beijing 100871, China; 3. State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100871, China;
Yi Chuan. 2016 May;38(5):391-401. doi: 10.16288/j.yczz.15-329.
High-throughput screening, a powerful tool for the discovery of functionally important genes responsible for certain phenotypes, is performed according to loss-of-function or gain-of-function strategies. RNAi technology or knockout approaches have been widely used in high throughput screening due to their advantages of ease use, low cost and so on. However, imcomplete knockdown activity and off-target effect hindered their utility. More recently, CRISPR/Cas9 technology is becoming a robust tool for genome editing in diverse cells or animals, since it could generate a gene mutation in a target-specific manner. In this review, we first summarize the characterization of CRISPR/Cas9 and make comparison with traditional genetic tools, then describe recent achievements of genetic screen in several model organisms using CRISPR/Cas9, finally discuss on its future challenges and opportunities.
高通量筛选是发现导致特定表型的功能重要基因的有力工具,它是根据功能丧失或功能获得策略进行的。RNAi技术或基因敲除方法因其易于使用、成本低等优点而被广泛应用于高通量筛选。然而,不完全的敲低活性和脱靶效应阻碍了它们的应用。最近,CRISPR/Cas9技术正成为一种用于在各种细胞或动物中进行基因组编辑的强大工具,因为它可以以靶标特异性方式产生基因突变。在本综述中,我们首先总结CRISPR/Cas9的特性并与传统遗传工具进行比较,然后描述使用CRISPR/Cas9在几种模式生物中进行遗传筛选的最新成果,最后讨论其未来的挑战和机遇。