基于CRISPR/Cas9系统高通量筛选研究功能基因

doi:10.16288/j.yczz.15-329

遗传 ›› 2016, Vol. 38 ›› Issue (5): 391-401.doi: 10.16288/j.yczz.15-329

基于CRISPR/Cas9系统高通量筛选研究功能基因

王干诚¹, 马明¹, 叶延帧¹, 席建忠^{1, 2, 3}

1. 北京大学工学院生物医学工程系,北京 100871;
2. 北京大学生物膜和膜技术国家重点实验室,分子医学研究所,北京 100871;
3. 北京大学天然药物及仿生药物国家重点实验室,北京 100871

收稿日期:2015-10-25 修回日期:2016-01-21 出版日期:2016-05-20 发布日期:2016-02-24
通讯作者: 席建忠,博士,教授,研究方向：生物医学工程。Tel: 010-62760698; E-mail: jzxi@pku.edu.cn E-mail:wgcd@pku.edu.cn
作者简介:王干诚,硕士研究生,专业方向：生物医学工程。E-mail: wgcd@pku.edu.cn

High-throughput functional screening using CRISPR/Cas9 system

Gancheng Wang¹, MingMa¹, Yanzhen Ye¹, Jianzhong Xi^{1, 2, 3}

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

Received:2015-10-25 Revised:2016-01-21 Online:2016-05-20 Published:2016-02-24

摘要/Abstract

摘要： 利用功能缺失型(Loss-of-function)或者功能获得型(Gain-of-function) 策略高通量筛选功能基因,是研究人员快速寻找调控特定表型的重要或关键基因的主要方法。RNA干扰(RNA interference,RNAi)的遗传筛选方法因操作简单、成本相对较低等优势,尽管已经得到了广泛的应用,然而其抑制效果不完全、脱靶效应明显等劣势依然存在。近年来兴起的CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeat sequences/ CRISPR-associated protein 9)技术能快速、简便、准确地实现基因组敲除等编辑功能,因而成为一种强大的遗传筛选工具;在各种细胞系、人和小鼠及斑马鱼等多种模式动物中,大规模运用该方法筛选功能基因已经取得了巨大成功。本文总结了CRISPR/Cas9技术的特点,将其与传统基因工程方法进行了分析比较,回顾了近期相关的高通量功能基因筛选工作,最后探讨了该技术未来的发展趋势。

关键词: CRISPR/Cas9, 基因组编辑技术, 高通量, 遗传筛选

Abstract: 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.

Key words: CRISPR/Cas9, genome editing technology, high-throughput, genetic screening

王干诚, 马明, 叶延帧, 席建忠. 基于CRISPR/Cas9系统高通量筛选研究功能基因[J]. 遗传, 2016, 38(5): 391-401.

Gancheng Wang, MingMa, Yanzhen Ye, Jianzhong Xi. High-throughput functional screening using CRISPR/Cas9 system[J]. HEREDITAS(Beijing), 2016, 38(5): 391-401.

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基于CRISPR/Cas9系统高通量筛选研究功能基因

High-throughput functional screening using CRISPR/Cas9 system

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