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Engineering of a promoter repressed by a light-regulated transcription factor in escherichia coli
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Camsund, Daniel, Jaramillo, Alfonso and Lindblad, Peter (2021) Engineering of a promoter repressed by a light-regulated transcription factor in escherichia coli. BioDesign Research, 2021 (9857418). doi:10.34133/2021/9857418 ISSN 2693-1257.
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WRAP-Engineering-promoter-repressed-light-regulated-transcription-escherichia-coli-22.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (783Kb) | Preview |
Official URL: https://doi.org/10.34133/2021/9857418
Abstract
Light-regulated gene expression systems allow controlling gene expression in space and time with high accuracy. Contrary to previous synthetic light sensors that incorporate two-component systems which require localization at the plasma membrane, soluble one-component repression systems provide several advantageous characteristics. Firstly, they are soluble and able to diffuse across the cytoplasm. Secondly, they are smaller and of lower complexity, enabling less taxing expression and optimization of fewer parts. Thirdly, repression through steric hindrance is a widespread regulation mechanism that does not require specific interaction with host factors, potentially enabling implementation in different organisms. Herein, we present the design of the synthetic promoter PEL that in combination with the light-regulated dimer EL222 constitutes a one-component repression system. Inspired by previously engineered synthetic promoters and the Escherichia coli lacZYA promoter, we designed PEL with two EL222 operators positioned to hinder RNA polymerase binding when EL222 is bound. PEL is repressed by EL222 under conditions of white light with a light-regulated repression ratio of five. Further, alternating conditions of darkness and light in cycles as short as one hour showed that repression is reversible. The design of the PEL-EL222 system herein presented could aid the design and implementation of analogous one-component optogenetic repression systems. Finally, we compare the PEL-EL222 system with similar systems and suggest general improvements that could optimize and extend the functionality of EL222-based as well as other one-component repression systems.
Item Type: | Journal Article | ||||||||||||||||||||||||||||||
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Subjects: | Q Science > QH Natural history > QH426 Genetics Q Science > QR Microbiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||||||||||||||||||||||||
SWORD Depositor: | Library Publications Router | ||||||||||||||||||||||||||||||
Library of Congress Subject Headings (LCSH): | Gene expression, Escherichia coli -- Genetics, Genetic regulation | ||||||||||||||||||||||||||||||
Journal or Publication Title: | BioDesign Research | ||||||||||||||||||||||||||||||
Publisher: | American Association for the Advancement of Science (AAAS) | ||||||||||||||||||||||||||||||
ISSN: | 2693-1257 | ||||||||||||||||||||||||||||||
Official Date: | 28 September 2021 | ||||||||||||||||||||||||||||||
Dates: |
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Volume: | 2021 | ||||||||||||||||||||||||||||||
Number: | 9857418 | ||||||||||||||||||||||||||||||
Number of Pages: | 11 | ||||||||||||||||||||||||||||||
DOI: | 10.34133/2021/9857418 | ||||||||||||||||||||||||||||||
Status: | Peer Reviewed | ||||||||||||||||||||||||||||||
Publication Status: | Published | ||||||||||||||||||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||||||||||||||||||||
Copyright Holders: | Copyright © 2021 Daniel Camsund et al. Exclusive Licensee Nanjing Agricultural University. | ||||||||||||||||||||||||||||||
Date of first compliant deposit: | 24 October 2022 | ||||||||||||||||||||||||||||||
Date of first compliant Open Access: | 24 October 2022 | ||||||||||||||||||||||||||||||
RIOXX Funder/Project Grant: |
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