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Genetically engineered light sensors for control of bacterial gene expression
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Photochemistry and Molecular Science.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Photochemistry and Molecular Science.
2011 (English)In: Biotechnology Journal, ISSN 1860-6768, Vol. 6, no 7, 826-836 p.Article, review/survey (Refereed) Published
Abstract [en]

Light of different wavelengths can serve as a transient, noninvasive means of regulating gene expression for biotechnological purposes. Implementation of advanced gene regulatory circuits will require orthogonal transcriptional systems that can be simultaneously controlled and that can produce several different control states. Fully genetically encoded light sensors take advantage of the favorable characteristics of light, do not need the supplementation of any chemical inducers or co-factors, and have been demonstrated to control gene expression in Escherichia coli. Herein, we review engineered light-sensor systems with potential for in vivo regulation of gene expression in bacteria, and highlight different means of extending the range of available light input and transcriptional output signals. Furthermore, we discuss advances in multiplexing different light sensors for achieving multichromatic control of gene expression and indicate developments that could facilitate the construction of efficient systems for light-regulated, multistate control of gene expression.

Place, publisher, year, edition, pages
2011. Vol. 6, no 7, 826-836 p.
Keyword [en]
Multichromatic, light-regulated promoter, Photoreceptor, Protein engineering, Synthetic biology
National Category
Chemical Sciences
URN: urn:nbn:se:uu:diva-156829DOI: 10.1002/biot.201100091ISI: 000292738100007OAI: oai:DiVA.org:uu-156829DiVA: diva2:433907
Available from: 2011-08-11 Created: 2011-08-09 Last updated: 2014-06-30Bibliographically approved
In thesis
1. Engineering Transcriptional Systems for Cyanobacterial Biotechnology
Open this publication in new window or tab >>Engineering Transcriptional Systems for Cyanobacterial Biotechnology
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Cyanobacteria are solar-powered cell factories that can be engineered to supply us with renewable fuels and chemicals. To do so robust and well-working biological parts and tools are necessary. Parts for controlling gene expression are of special importance in living systems, and specifically promoters are needed for enabling and simplifying rational design. Synthetic biology is an engineering science that incorporates principles such as decoupling, standardization and modularity to enable the design and construction of more advanced systems from simpler parts and the re-use of parts in new contexts. For these principles to work, cross-talk must be avoided and therefore orthogonal parts and systems are important as they are decoupled by definition. This work concerns the design and development of biological parts and tools that can enable synthetic biology in cyanobacteria. This encompasses parts necessary for the development of other systems, such as vectors and translational elements, but with a focus on transcriptional regulation. First, to enable the development and characterization of promoters in different cyanobacterial chassis, a broad-host-range BioBrick plasmid, pPMQAK1, was constructed and confirmed to function in several cyanobacterial strains. Then, ribosome binding sites, protease degradation tags and constitutive, orthogonal promoters were characterized in the model strain Synechocystis PCC 6803. These tools were then used to design LacI-regulated promoter libraries for studying DNA-looping and the behaviour of LacI-mediated loops in Synechocystis. Ultimately, this lead to the design of completely repressed LacI-regulated promoters that could be used for e.g. cyanobacterial genetic switches, and was used to design a destabilized version of the repressed promoter that could be induced to higher levels. Further, this promoter was used to implement an orthogonal transcriptional system based on T7 RNAP that was shown to drive different levels of T7 promoter transcription depending on regulation. Also, Gal4-repressed promoters for bacteria were engineered and examined in Escherichia coli as an initial step towards transferring them to cyanobacteria. Attempts were also made to implement a light-regulated one-component transcription factor based on Gal4. This work provides a background for engineering transcription and provides suggestions for how to develop the parts further.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. 63 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1149
Cyanobacteria, Synthetic biology, promoters, transcription, LacI, Gal4, Light-regulation
National Category
Biochemistry and Molecular Biology
Research subject
Chemistry with specialization in Microbial Chemistry
urn:nbn:se:uu:diva-223599 (URN)978-91-554-8954-0 (ISBN)
Public defence
2014-06-05, Häggsalen, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:15 (English)
Available from: 2014-05-15 Created: 2014-04-22 Last updated: 2014-06-30

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Camsund, DanielLindblad, Peter
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