CRISPR系统及其应用于小鼠的研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2017-0530

摘要/Abstract

摘要： CRISPR系统作为一种基因编辑工具,自2013年首次应用于哺乳动物以来,凭借其设计简单,耗时短,效率高的优点,在短短四年间飞速发展,其应用已经远远超过了锌指核酸酶和转录激活因子样效应物核酸酶技术,并在分子生物学领域得到广泛应用。基于该系统发展而来的各项技术不断的被发掘,小鼠作为一种基本的实验动物,在CRISPR系统应用中扮演重要角色。目前基于CRISPR系统发展而来的应用工具已经成功的作用于各种体外培养的细胞,通过对胚胎细胞的编辑而产生动物模型的技术也在不断成熟,以及在疾病治疗特别是遗传病治疗中也展现出其独有的优势,并且其作用范围在不断的扩展,作用方式还在不断的创新。主要介绍CRISPR系统的结构与作用原理,讲述基因敲入、基因敲除、基因沉默、转录调控等作用于DNA的几种主要的CRISPR应用工具,针对目前主要的缺点即脱靶效应所研究出的几个改善措施,以及在小鼠疾病模型建立与疾病治疗中发挥的重要作用。

关键词: CRISPR系统, 基因调控, 脱靶效应, 小鼠

Abstract: As a gene editing tool,CRISPR system has been in rapid development since the first application in mammal in 2013, and iIt has been widely used in the field of molecular biology with its simple design,short time consuming,and high efficiency advantages. So far,its applications have far exceeded ZFNs and TALEN technology. With the continuous exploration of varied techniques from the CRISPR system,mice as a basic experimental animal play an important role in the application of CRISPR system. At present,the application tools developed from CRISPR system have been successfully applied to a variety of cells cultured in vitro. The technology for producing animal models by editing embryonic cells is also maturing,and it has its unique advantages in the treatment of diseases,especially in the treatment of hereditary diseases. And its scope of application is still expanding,and the mode of application is also in constant innovation. This paper mainly introduces the structure and mechanism of CRISPR system;also describes several major CRISPR application tools acting on DNA for gene knockin,knockout,gene silencing,transcriptional regulation,etc.,discuss several improvement measures focusing on its major deficiency,i.e.,the off-target effects,and its important role in the establishment of mouse disease models and treatment of diseases.

Key words: CRISPR/Cas9 system, gene regulation, off-target effects, mice

董维鹏,王君实,张少华,燕炯. CRISPR系统及其应用于小鼠的研究进展[J]. 生物技术通报, 2018, 34(5): 57-63.

DONG Wei-peng, WANG Jun-shi, ZHANG Shao-hua, YAN Jiong. Research Progress of CRISPR System and Its Application in Mice[J]. Biotechnology Bulletin, 2018, 34(5): 57-63.

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