Design of an intracellular aptamer-based fluorescent biosensor to track burden in Escherichia coli
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
Cell burden impacts the performance of engineered genetic constructs with great interest towards the development of tools to track it and improve biotechnology applications. Fluorogenic RNA aptamers are excellent candidates for live monitoring of burden because their production is expected to impose negligible load on the host. Here, we characterised a library of aptamers when expressed from different promoters in two Escherichia coli strains. We found that aptamer performance is dependent on the context of expression, and that, contrary to expectation, aptamer production impacts host fitness. We then built a library of burden-responsive biosensors testing their response to heterologous expression. The tRNA-Broc biosensor was selected for its fluorescence response, minimal impact on growth and ability to differentiate the burden imposed by different expression levels and constructs. The biosensor developed here adds to the collection of tools available to characterise burden and support applications where improved host performance is sought.
Date Issued
2025-10-01
Date Acceptance
2025-06-04
Citation
Trends in Biotechnology, 2025, 43 (10), pp.2566-2585
ISSN
0167-7799
Publisher
Cell Press
Start Page
2566
End Page
2585
Journal / Book Title
Trends in Biotechnology
Volume
43
Issue
10
Copyright Statement
© 2025 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/40610258
PII: S0167-7799(25)00222-7
Subjects
Biotechnology & Applied Microbiology
GENE-EXPRESSION
GROWTH
Life Sciences & Biomedicine
PROTEIN
REVEALS
RNA
Science & Technology
TRANSCRIPTION
Publication Status
Published
Coverage Spatial
England
Date Publish Online
2025-07-02