title: Biosensing Vibrio cholerae with Genetically Engineered Escherichia coli.
abstract: Cholera is a potentially mortal, infectious disease caused by Vibrio cholerae bacterium. Current treatment methods of cholera still have limitations. Beneficial microbes that could sense and kill the V. cholerae could offer potential alternative to preventing and treating cholera. However, such V. cholerae targeting microbe is still not available. This microbe requires a sensing system to be able to detect the presence of V. cholera bacterium. To this end, we designed and created a synthetic genetic sensing system using nonpathogenic Escherichia coli as the host. To achieve the system, we have moved proteins used by V. cholerae for quorum sensing into E. coli. These sensor proteins have been further layered with a genetic inverter based on CRISPRi technology. Our design process was aided by computer models simulating in vivo behavior of the system. Our sensor shows high sensitivity to presence of V. cholerae supernatant with tight control of expression of output GFP protein.
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore 639798.
authors.forename: Maciej B
authors.initials: MB
authors.lastname: Holowko
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore 639798.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore 639798.
authors.forename: Premkumar
authors.initials: P
authors.lastname: Jayaraman
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University , Singapore 639798.
authors.forename: Chueh Loo
authors.initials: CL
authors.lastname: Poh
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D015374:Biosensing Techniques; D064113:CRISPR-Cas Systems; D003198:Computer Simulation; D004926:Escherichia coli / Q000235:genetics*; D000074041:Microorganisms, Genetically-Modified; D011401:Promoter Regions, Genetic; D053038:Quorum Sensing; D017422:Sequence Analysis, DNA; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins
keywords: CRISPRi; Vibrio cholerae; biosensor; cholera; synthetic biology