Burden-driven feedback control of gene expression

Francesca Ceroni, Alice Boo, Simone Furini, Thomas E. Gorochowski, Olivier Borkowski, Yaseen N. Ladak, Ali R. Awan, Charlie Gilbert, Guy Bart Stan*, Tom Ellis

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

76 Citations (Scopus)
457 Downloads (Pure)

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.

Original languageEnglish
Pages (from-to)387-393
Number of pages7
JournalNature Methods
Volume15
Issue number5
Early online date26 Mar 2018
DOIs
Publication statusPublished - 1 May 2018

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute

Keywords

  • synthetic biology
  • feedback control
  • genetic circuits
  • metabolic engineering
  • biotechnology
  • systems biology

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