细菌中CRISPR/Cas系统的应用和优化

Application and optimization of CRISPR/Cas system in bacteria

摘要:

成簇规律间隔短回文重复序列 (Clustered regularly interspaced short palindromic repeats，CRISPR) 系统在近年来得到越来越广泛的应用。相比传统的基因组编辑技术，CRISPR/Cas系统具有编辑效率高、特异性强、花费低、对实验技术要求低等优势。其中Ⅱ型和Ⅴ型CRISPR/Cas系统分别仅需要单个Cas9蛋白和单个Cpf1蛋白作为切割双链DNA的工具，因此特别受到研究者们的青睐。目前CRISPR/Cas9技术已成功应用于斑马鱼、小鼠、人类细胞等真核生物的基因组编辑中，并取得一系列重要成果，但在细菌领域进行的相关研究并不多。文中将简单描述CRISPR/Cas系统及其作用机制，重点介绍该系统的优化以及近年来其在细菌学领域所取得的 进展。

Abstract:

Clustered regular interspaced short palindromic repeats (CRISPR) system has been widely used in recent years. Compared with traditional genome editing technology, CRISPR/Cas system has notable advantages, including high editing efficiency, high specificity, low cost and the convenience for manipulation. Type Ⅱ and Ⅴ CRISPR/Cas system only requires a single Cas9 protein or a single Cpf1 protein as effector nucleases for cutting double-stranded DNA, developed as genome editing tools. At present, CRISPR/Cas9 technology has been successfully applied to the genome editing of eukaryotes such as zebrafish, mice and human cells, whereas limited progress has been made in the genome editing of bacteria. In our review, we describe CRISPR/Cas system, its mechanism and summarize the optimization and progress of genome editing in bacteria.