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Cited by 75RNA and protein synthesis mechanismsBacterial Genetics and BiotechnologyMicrobial Metabolic Engineering and Bioproduction

Modular riboswitch toolsets for synthetic genetic control in diverse bacterial species

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Robinson, Christopher J., Vincent, Helen A., Wu, Ming-Cheng, Lowe, Phillip T., Dunstan, Mark S., Leys, David, Micklefield, Jason·2014·Journal of the American Chemical Society
DOI 10.1021/ja502873j
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Abstract

Ligand-dependent control of gene expression is essential for gene functional analysis, target validation, protein production, and metabolic engineering. However, the expression tools currently available are difficult to transfer between species and exhibit limited mechanistic diversity. Here we demonstrate how the modular architecture of purine riboswitches can be exploited to develop orthogonal and chimeric switches that are transferable across diverse bacterial species, modulating either transcription or translation, to provide tunable activation or repression of target gene expression, in response to synthetic non-natural effector molecules. Our novel riboswitch–ligand pairings are shown to regulate physiologically important genes required for bacterial motility in Escherichia coli and cell morphology in Bacillus subtilis. These findings are relevant for future gene function studies and antimicrobial target validation, while providing new modular and orthogonal regulatory components for deployment in synthetic biology regimes.

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  • The riboswitch sequence in RiboTite is a modified version of the adenine-sensing A-riboswitch from Vibrio vulnificus and is controlled by pyrimido-pyrimidine-2,4-diamine, so expression of the target gene occurs only in the presence of both IPTG and PPDA, reducing leaky expression to almost undetectable levels.

    “The riboswitch sequence, a modified version of the adenine-sensing A-riboswitch from Vibrio vulnificus, is controlled by pyrimido-pyrimidine-2,4-diamine (PPDA) [178], thereby the expression of the target gene occurs only in the presence of both IPTG and PPDA, which effectively reduces the leaky expression to almost undetectable levels.”

    From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [178]

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Cited by

Sorted by citations

Authors
Robinson, C. J., Vincent, H. A., Wu, M.-C., Lowe, P. T., Dunstan, M. S., Leys, D., & Micklefield, J.
Affiliations
University of Manchester (GB)
Corresponding author
Micklefield, Jason
ORCID
Venue
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Published
2014-06-27
Volume
136
Issue
30
Pages
10615-10624
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
DOI
10.1021/ja502873j
OpenAlex
W2085802860
Cited by
75
References
0
Citations by year
2026: 3 · 2024: 4 · 2023: 4 · 2022: 2 · 2021: 6 · 2020: 7 · 2019: 11 · 2018: 11 · 2017: 8 · 2016: 7 · 2015: 9 · 2014: 2 · 2013: 1
FWCI
3.55
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of the American Chemical Society (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research Explorer (The University of Manchester) (submittedVersion, cc-by)
Funders
Biotechnology and Biological Sciences Research Council (BB/I012648/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
purine riboswitch, transcriptional regulation, translational regulation, synthetic biology toolkit, bacterial motility, cell morphology, Escherichia coli, Bacillus subtilis
MeSH terms
Bacillus subtilis, Base Sequence, Escherichia coli, Models, Molecular, Molecular Sequence Data, Gene Expression Regulation, Bacterial, Aptamers, Nucleotide, Riboswitch

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