Burden-driven feedback control of gene expression
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
Cells use feedback regulation to ensure robust growth despite fluctuating demands for resources and differing environmental conditions. However, the expression of foreign proteins from engineered constructs is an unnatural burden that cells are not adapted for. Here we combined RNA-seq with an in vivo assay to identify the major transcriptional changes that occur in Escherichia coli when inducible synthetic constructs are expressed. We observed that native promoters related to the heat-shock response activated expression rapidly in response to synthetic expression, regardless of the construct. Using these promoters, we built a dCas9-based feedback-regulation system that automatically adjusts the expression of a synthetic construct in response to burden. Cells equipped with this general-use controller maintained their capacity for native gene expression to ensure robust growth and thus outperformed unregulated cells in terms of protein yield in batch production. This engineered feedback is to our knowledge the first example of a universal, burden-based biomolecular control system and is modular, tunable and portable.
Date Issued
2018-05-01
Date Acceptance
2018-02-01
Citation
Nature Methods, 2018, 15 (5), pp.387-393
ISSN
1548-7091
Publisher
Nature Research
Start Page
387
End Page
393
Journal / Book Title
Nature Methods
Volume
15
Issue
5
Copyright Statement
© 2018 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Engineering & Physical Science Research Council (EPSRC)
Engineering & Physical Science Research Council (EPSRC)
Engineering & Physical Science Research Council (EPSRC)
Identifier
https://www.nature.com/articles/nmeth.4635
Grant Number
EP/J021849/1
EP/M002306/1
EP/M002187/1
EP/P009352/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
HEAT-SHOCK RESPONSE
RNA-SEQ
TRADE-OFFS
VISUALIZATION
SIGMA(32)
PROTEINS
DYNAMICS
BACTERIA
REVEALS
DESIGNS
Escherichia coli
Gene Expression Regulation, Bacterial
High-Throughput Nucleotide Sequencing
Plasmids
Promoter Regions, Genetic
Sequence Analysis, RNA
Synthetic Biology
Transcription, Genetic
Escherichia coli
Sequence Analysis, RNA
Transcription, Genetic
Gene Expression Regulation, Bacterial
Plasmids
Promoter Regions, Genetic
Synthetic Biology
High-Throughput Nucleotide Sequencing
Developmental Biology
06 Biological Sciences
10 Technology
11 Medical and Health Sciences
Publication Status
Published
Date Publish Online
2018-03-26