Synthetic negative feedback circuits using engineered small RNAs
Author(s)
Type
Journal Article
Abstract
Negative feedback is known to enable biological and man-made systems to perform reliably in the face of uncertainties and disturbances. To date, synthetic biological feedback circuits have primarily relied upon protein-based, transcriptional regulation to control circuit output. Small RNAs (sRNAs) are non-coding RNA molecules that can inhibit translation of target messenger RNAs (mRNAs). In this work, we modelled, built and validated two synthetic negative feedback circuits that use rationally-designed sRNAs for the first time. The first circuit builds upon the well characterised tet-based autorepressor, incorporating an externally-inducible sRNA to tune the effective feedback strength. This allows more precise fine-tuning of the circuit output in contrast to the sigmoidal, steep input-output response of the autorepressor alone. In the second circuit, the output is a transcription factor that induces expression of an sRNA, which inhibits translation of the mRNA encoding the output, creating direct, closed-loop, negative feedback. Analysis of the noise profiles of both circuits showed that the use of sRNAs did not result in large increases in noise. Stochastic and deterministic modelling of both circuits agreed well with experimental data. Finally, simulations using fitted parameters allowed dynamic attributes of each circuit such as response time and disturbance rejection to be investigated.
Date Issued
2018-10-12
Date Acceptance
2018-09-06
Citation
Nucleic Acids Research, 2018, 46 (18), pp.9875-9889
ISSN
0305-1048
Publisher
Oxford University Press
Start Page
9875
End Page
9889
Journal / Book Title
Nucleic Acids Research
Volume
46
Issue
18
Copyright Statement
©
The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http:
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creativecommons.org
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licenses
/
by
/
4.0
/
), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http:
//
creativecommons.org
/
licenses
/
by
/
4.0
/
), which
permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/30212900
PII: 5096075
Grant Number
BB/M011321/1
Subjects
05 Environmental Sciences
06 Biological Sciences
08 Information And Computing Sciences
Developmental Biology
Publication Status
Published
Coverage Spatial
England
Date Publish Online
2018-09-13