title: Genome engineering and gene expression control for bacterial strain development.
abstract: In recent years, a number of techniques and tools have been developed for genome engineering and gene expression control to achieve desired phenotypes of various bacteria. Here we review and discuss the recent advances in bacterial genome manipulation and gene expression control techniques, and their actual uses with accompanying examples. Genome engineering has been commonly performed based on homologous recombination. During such genome manipulation, the counterselection systems employing SacB or nucleases have mainly been used for the efficient selection of desired engineered strains. The recombineering technology enables simple and more rapid manipulation of the bacterial genome. The group II intron-mediated genome engineering technology is another option for some bacteria that are difficult to be engineered by homologous recombination. Due to the increasing demands on high-throughput screening of bacterial strains having the desired phenotypes, several multiplex genome engineering techniques have recently been developed and validated in some bacteria. Another approach to achieve desired bacterial phenotypes is the repression of target gene expression without the modification of genome sequences. This can be performed by expressing antisense RNA, small regulatory RNA, or CRISPR RNA to repress target gene expression at the transcriptional or translational level. All of these techniques allow efficient and rapid development and screening of bacterial strains having desired phenotypes, and more advanced techniques are expected to be seen.
authors.affiliation: 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, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea; BioProcess Engineering Research Center, KAIST, Daejeon, Republic of Korea.
authors.forename: Chan Woo
authors.initials: CW
authors.lastname: Song
authors.forename: Joungmin
authors.initials: J
authors.lastname: Lee
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
mesh_terms: D001419:Bacteria / Q000235:genetics*; D064113:CRISPR-Cas Systems / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial / Q000235:genetics*; D060847:Metabolic Engineering; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism
chemical_list: D012329:RNA, Bacterial
keywords: Bacteria; CRISPR-Cas9; Genome engineering; Metabolic engineering; Small regulatory RNA