title: CRISPR technologies for bacterial systems: Current achievements and future directions.
abstract: Throughout the decades of its history, the advances in bacteria-based bio-industries have coincided with great leaps in strain engineering technologies. Recently unveiled clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) systems are now revolutionizing biotechnology as well as biology. Diverse technologies have been derived from CRISPR/Cas systems in bacteria, yet the applications unfortunately have not been actively employed in bacteria as extensively as in eukaryotic organisms. A recent trend of engineering less explored strains in industrial microbiology-metabolic engineering, synthetic biology, and other related disciplines-is demanding facile yet robust tools, and various CRISPR technologies have potential to cater to the demands. Here, we briefly review the science in CRISPR/Cas systems and the milestone inventions that enabled numerous CRISPR technologies. Next, we describe CRISPR/Cas-derived technologies for bacterial strain development, including genome editing and gene expression regulation applications. Then, other CRISPR technologies possessing great potential for industrial applications are described, including typing and tracking of bacterial strains, virome identification, vaccination of bacteria, and advanced antimicrobial approaches. For each application, we note our suggestions for additional improvements as well. In the same context, replication of CRISPR/Cas-based chromosome imaging technologies developed originally in eukaryotic systems is introduced with its potential impact on studying bacterial chromosomal dynamics. Also, the current patent status of CRISPR technologies is reviewed. Finally, we provide some insights to the future of CRISPR technologies for bacterial systems by proposing complementary techniques to be developed for the use of CRISPR technologies in even wider range of applications.
authors: Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejeon 34141, Republic of Korea. Electronic address: chkr1018@kaist.ac.kr.
authors: Kyeong Rok
authors: KR
authors: Choi
authors: Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejeon 34141, Republic of Korea; BioProcess Engineering Research Center, KAIST, Daejeon 34141, Republic of Korea; BioInformatics Research Center, KAIST, Daejeon 34141, Republic of Korea; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Alle 6, Hørsholm 2970, Denmark. Electronic address: leesy@kasist.ac.kr.
authors: Sang Yup
authors: SY
authors: Lee
mesh_terms: D001419:Bacteria / Q000235:genetics*; D001431:Bacteriological Techniques / Q000639:trends*; D064130:CRISPR-Associated Proteins / Q000235:genetics; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D000072669:Gene Editing / Q000639:trends; D024861:Genetic Enhancement
chemical_list: D064130:CRISPR-Associated Proteins
keywords: Antimicrobials; CRISPR/Cas system; Cas9, bacteria; Expression regulation; Genome editing; Imaging; Typing; Vaccination of bacteria; Virome