Anton Tobias, Karg Elisabeth, Bultmann Sebastian
Department of Biology II and Center for Integrated Protein Science Munich (CIPSM), LMU Munich, 82152 Martinsried, Germany.
Biol Methods Protoc. 2018 May 10;3(1):bpy002. doi: 10.1093/biomethods/bpy002. eCollection 2018.
Since the discovery of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas) as a tool for gene editing a plethora of locus-specific as well as genome-wide approaches have been developed that allow efficient and reproducible manipulation of genomic sequences. However, the seemingly unbound potential of CRISPR/Cas does not stop with its utilization as a site-directed nuclease. Mutations in its catalytic centers render Cas9 (dCas9) a universal recruitment platform that can be utilized to control transcription, visualize DNA sequences, investigate proteome compositions and manipulate epigenetic modifications at user-defined genomic loci. In this review, we give a comprehensive introduction and overview of the development, improvement and application of recent dCas9-based approaches.
自从成簇规律间隔短回文重复序列(CRISPR)/CRISPR相关系统(Cas)作为一种基因编辑工具被发现以来,已经开发出了大量位点特异性以及全基因组范围的方法,这些方法能够对基因组序列进行高效且可重复的操作。然而,CRISPR/Cas看似无限的潜力并不局限于将其用作定点核酸酶。其催化中心的突变使Cas9(dCas9)成为一个通用的募集平台,可用于控制转录、可视化DNA序列、研究蛋白质组组成以及在用户定义的基因组位点操纵表观遗传修饰。在本综述中,我们对基于dCas9的最新方法的开发、改进及应用进行了全面的介绍和概述。