Fundamental CRISPR-Cas9 tools and current applications in microbial systems

Synthetic and Systems Biotechnology

Yajun Yan¹  Justin Forrest Rey¹  Xiaolin Shen²  Qipeng Yuan²  Jia Wang²  Pingfang Tian² 

[1] College of Engineering, The University of Georgia, Athens, GA 30602, USA;College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China;

关键词: CRISPR-Cas9;    CRISPR interference;    CRISPR activation;    DNA repair;    Homologous recombination;   

DOI  :  10.1016/j.synbio.2017.08.006

来源: DOAJ

【 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.

【 授权许可】

Unknown