CRISPR/Cas基因编辑系统在原核微生物细胞工厂构建中的开发与应用

Development and application of CRISPR/Cas genome editing system in the construction of prokaryotic microbial cell factories

摘要:

随着能源和环境问题的日益突出，化学品以及燃料的合成方式正逐渐由传统的化学法合成转变为以细菌为基础的生物炼制过程，其中最关键问题是需要开发出合适的基因工程工具用于构建相应的产品生产菌株。成簇的规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats，CRISPR)/CRISPR相关蛋白(CRISPR-associated proteins，Cas)系统是一种存在于细菌和古细菌中的免疫系统，能够用于抵御病毒和外源质粒的入侵，近年来被开发成为一种高效、便捷、精确的基因编辑工具，显示出巨大的应用潜力。本文立足于CRISPR/Cas系统的原理与最新分类，结合实例综述了CRISPR/Cas基因编辑系统在原核微生物细胞工厂构建中的建立与优化策略，以及主要的应用方向，并探讨该系统所面临的主要问题并提出了一些可行的解决方案。

Abstract:

As the energy crisis and environmental problems are ongoing and getting worse, traditional chemical synthesis industry is being replaced by bacteria-based bio-industries for production of chemicals and fuels, in which, the most critical issue is developing proper genetic engineering tools to establish platform organisms for bioproduction. Derived from archaeal and prokaryotic immune systems to defend viral and plasmid invasions, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) are developed into a fast, easy, and robust genome editing tool in recent years and have potential to cater to the demands. Here, we briefly review the principle and advanced classification of CRISPR/Cas systems. The establishment, optimization and applications of CRISPR-based genome editing tools in the construction of prokaryotic microbial cell factories are introduced with the examples. Besides, some feasible solutions are proposed based on the discussion about the problems existed in CRISPR/Cas systems.