Refactoring and Optimization of Light-Switchable Escherichia coli Two-Component Systems

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dc.citation.firstpage820en_US
dc.citation.issueNumber11en_US
dc.citation.journalTitleACS Synthetic Biologyen_US
dc.citation.lastpage831en_US
dc.citation.volumeNumber3en_US
dc.contributor.authorSchmidl, Sebastian R.en_US
dc.contributor.authorSheth, Ravi U.en_US
dc.contributor.authorWu, Andrewen_US
dc.contributor.authorTabor, Jeffrey J.en_US
dc.contributor.orgBioengineeringen_US
dc.contributor.orgBiosciencesen_US
dc.date.accessioned2016-02-05T18:38:32Zen_US
dc.date.available2016-02-05T18:38:32Zen_US
dc.date.issued2014en_US
dc.description.abstractLight-switchable proteins enable unparalleled control of molecular biological processes in live organisms. Previously, we have engineered red/far-red and green/red photoreversible two-component signal transduction systems (TCSs) with transcriptional outputs in E. coli and used them to characterize and control synthetic gene circuits with exceptional quantitative, temporal, and spatial precision. However, the broad utility of these light sensors is limited by bulky DNA encoding, incompatibility with commonly used ligand-responsive transcription factors, leaky output in deactivating light, and less than 10-fold dynamic range. Here, we compress the four genes required for each TCS onto two streamlined plasmids and replace all chemically inducible and evolved promoters with constitutive, engineered versions. Additionally, we systematically optimize the expression of each sensor histidine kinase and response regulator, and redesign both pathway output promoters, resulting in low leakiness and 72- and 117-fold dynamic range, respectively. These second-generation light sensors can be used to program the expression of more genes over a wider range and can be more easily combined with additional plasmids or moved to different host strains. This work demonstrates that bacterial TCSs can be optimized to function as high-performance sensors for scientific and engineering applications.en_US
dc.identifier.citationSchmidl, Sebastian R., Sheth, Ravi U., Wu, Andrew, et al.. "Refactoring and Optimization of Light-Switchable Escherichia coli Two-Component Systems." <i>ACS Synthetic Biology,</i> 3, no. 11 (2014) American Chemical Society: 820-831. http://dx.doi.org/10.1021/sb500273n.en_US
dc.identifier.doihttp://dx.doi.org/10.1021/sb500273nen_US
dc.identifier.urihttps://hdl.handle.net/1911/88397en_US
dc.language.isoengen_US
dc.publisherAmerican Chemical Societyen_US
dc.rightsThis is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.en_US
dc.rights.urihttp://pubs.acs.org/page/policy/authorchoice_termsofuse.htmlen_US
dc.subject.keywordoptogeneticsen_US
dc.subject.keywordEscherichia colien_US
dc.subject.keywordtwo component systemen_US
dc.subject.keywordphytochromeen_US
dc.subject.keywordcyanobacteriochromeen_US
dc.subject.keywordrefactoringen_US
dc.titleRefactoring and Optimization of Light-Switchable Escherichia coli Two-Component Systemsen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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