Tian Pingfang, Wang Jia, Shen Xiaolin, Rey Justin Forrest, Yuan Qipeng, Yan Yajun
College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Synth Syst Biotechnol. 2017 Sep 8;2(3):219-225. doi: 10.1016/j.synbio.2017.08.006. eCollection 2017 Sep.
Derived from the bacterial adaptive immune system, CRISPR technology has revolutionized conventional genetic engineering methods and unprecedentedly facilitated strain engineering. In this review, we outline the fundamental CRISPR tools that have been employed for strain optimization. These tools include CRISPR editing, CRISPR interference, CRISPR activation and protein imaging. To further characterize the CRISPR technology, we present current applications of these tools in microbial systems, including model- and non-model industrial microorganisms. Specially, we point out the major challenges of the CRISPR tools when utilized for multiplex genome editing and sophisticated expression regulation. To address these challenges, we came up with strategies that place emphasis on the amelioration of DNA repair efficiency through CRISPR-Cas9-assisted recombineering. Lastly, multiple promising research directions were proposed, mainly focusing on CRISPR-based construction of microbial ecosystems toward high production of desired chemicals.
源自细菌适应性免疫系统的CRISPR技术彻底改变了传统基因工程方法，并前所未有的促进了菌株工程。在本综述中，我们概述了用于菌株优化的基本CRISPR工具。这些工具包括CRISPR编辑、CRISPR干扰、CRISPR激活和蛋白质成像。为了进一步表征CRISPR技术，我们介绍了这些工具在微生物系统中的当前应用，包括模式和非模式工业微生物。特别地，我们指出了CRISPR工具在用于多重基因组编辑和复杂表达调控时的主要挑战。为应对这些挑战，我们提出了强调通过CRISPR-Cas9辅助重组工程提高DNA修复效率的策略。最后，提出了多个有前景的研究方向，主要集中在基于CRISPR构建微生物生态系统以高产所需化学品。
