title: Riboselector: riboswitch-based synthetic selection device to expedite evolution of metabolite-producing microorganisms.
abstract: Many successful metabolic engineering projects have utilized evolutionary approaches, which consist of generating phenotypic diversity and screening for desired phenotype. Since conventional screening methods suffer from low throughput and limited target metabolites, a universal high-throughput screening platform for selection of improved strains should be developed to facilitate evolution of metabolite high producer. Recently, riboswitches have received attention as attractive sensor-actuator hybrids that can control gene expression in response to intracellular metabolite concentration. Our group developed a riboswitch-based selection device called "Riboselector" which can give a growth advantage to metabolite-overproducing strains by modulating expression of a selectable marker gene. We applied the device to expedite evolution of lysine producing Escherichia coli, and the selected strain showed a dramatic improvement of lysine production compared to its parental strain. Moreover, a tryptophan-responsive Riboselector was also developed using synthetic tryptophan aptamer. In this chapter, we provide a step-by-step overview of developing synthetic RNA devices comprising a riboswitch and a selection module that specifically sense inconspicuous metabolites and enrich high producer strains out of library.
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea.
authors.forename: Sungho
authors.initials: S
authors.lastname: Jang
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea.
authors.forename: Jina
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea.
authors.forename: Sang Woo
authors.initials: SW
authors.lastname: Seo
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea; School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Gyeongbuk, Republic of Korea. Electronic address: gyjung@postech.ac.kr.
authors.forename: Gyoo Yeol
authors.initials: GY
authors.lastname: Jung
mesh_terms: D015964:Gene Expression Regulation, Bacterial; D060847:Metabolic Engineering / Q000379:methods*; D058928:Riboswitch / Q000235:genetics*; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D058928:Riboswitch
keywords: Enrichment; Metabolic engineering; Metabolite production; Riboselector; Riboswitch; Screening device; Synthetic biology