Optogenetic Control of Bacterial Expression by Red Light

Abstract

In optogenetics, as in nature, sensory photoreceptors serve to control cellular processes by light. Bacteriophytochrome (BphP) photoreceptors sense red and far-red light via a biliverdin chromophore and, in response, cycle between the spectroscopically, structurally, and functionally distinct Pr and Pfr states. BphPs commonly belong to two-component systems that control the phosphorylation of cognate response regulators and downstream gene expression through histidine kinase modules. We recently demonstrated that the paradigm BphP from Deinococcus radiodurans exclusively acts as a phosphatase but that its photosensory module can control the histidine kinase activity of homologous receptors. Here, we apply this insight to reprogram two widely used setups for bacterial gene expression from blue-light to red-light control. The resultant pREDusk and pREDawn systems allow gene expression to be regulated down and up, respectively, uniformly under red light by 100-fold or more. Both setups are realized as portable, single plasmids that encode all necessary components including the biliverdin-producing machinery. The triggering by red light affords high spatial resolution down to the single-cell level. As pREDusk and pREDawn respond sensitively to red light, they support multiplexing with optogenetic systems sensitive to other light colors. Owing to the superior tissue penetration of red light, the pREDawn system can be triggered at therapeutically safe light intensities through material layers, replicating the optical properties of the skin and skull. Given these advantages, pREDusk and pREDawn enable red-light-regulated expression for diverse use cases in bacteria.

Publication DOI: https://doi.org/10.1021/acssynbio.2c00259
Divisions: College of Engineering & Physical Sciences > School of Engineering and Technology > Mechanical, Biomedical & Design
College of Engineering & Physical Sciences
Aston University (General)
Additional Information: © 2022, The Authors. Published by American Chemical Society with a Creative Commons Attribution (CC-BY) License [https://creativecommons.org/licenses/by/4.0/]. Funding Information: This work was supported by the Academy of Finland grant 330678 (H.T.), Three-year grant 2018–2020 from the University of Helsinki (E.M and H.T.), and Bayreuth Humboldt Centre Senior Fellowship 2020 (E. M., A.M., and H.T.). A.M. acknowledges support by the Deutsche Forschungsgemeinschaft (MO2192/6–2) and the European Commission (FET Open NEUROPA, grant agreement 863214). Biomedicum Imaging Unit (BIU) core facility, University of Helsinki is acknowledged for their microscopy services. HiLife Flow Cytometry Unit, University of Helsinki is acknowledged for their flow cytometry services.
Uncontrolled Keywords: gene expression,optogenetics,phytochrome,sensory photoreceptor,signal transduction,two-component system,Biomedical Engineering,Biochemistry, Genetics and Molecular Biology (miscellaneous)
Publication ISSN: 2161-5063
Last Modified: 02 Dec 2024 08:43
Date Deposited: 14 Sep 2022 15:32
Full Text Link:
Related URLs: https://pubs.ac ... ssynbio.2c00259 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2022-10-21
Published Online Date: 2022-08-23
Accepted Date: 2022-05-17
Authors: Multamäki, Elina
García de Fuentes, Andrés
Sieryi, Oleksii
Bykov, Alexander
Gerken, Uwe
Ranzani, Américo Tavares
Köhler, Jürgen
Meglinski, Igor (ORCID Profile 0000-0002-7613-8191)
Möglich, Andreas
Takala, Heikki

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