Fundamental CRISPR-Cas9 tools and current applications in microbial systems  

作　　者：Pingfang Tian Jia Wang Xiaolin Shen Justin Forrest Rey Qipeng Yuan Yajun Yan 

机构地区：[1]College of Life Science and Technology,Beijing University of Chemical Technology,Beijing 100029,China [2]State Key Laboratory of Chemical Resource Engineering,Beijing University of Chemical Technology,Beijing 100029,China [3]Beijing Advanced Innovation Center for Soft Matter Science and Engineering,Beijing University of Chemical Technology,Beijing 100029,China [4]College of Engineering,The University of Georgia,Athens,GA 30602,USA

出　　处：《Synthetic and Systems Biotechnology》2017年第3期219-225,共7页合成和系统生物技术（英文）

基　　金：This work was supported by grants from National Natural Science Foundation of China(No.21276014,21476011);National High Technology Research and Development Program(863 Program)(No.2015AA021003);Fundamental Research Funds for the Central Universities(YS1407);111 project(B13005).

摘　　要：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.

关 键 词：CRISPR-Cas9 CRISPR interference CRISPR activation DNA repair Homologous recombination 

分 类 号：Q78[生物学—分子生物学]