Engineering a Functional Small RNA Negative Autoregulation Network with Model-Guided Design

Chelsea Y. Hu, Melissa K. Takahashi, Yan Zhang, Julius B. Lucks*

*Corresponding author for this work

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

RNA regulators are powerful components of the synthetic biology toolbox. Here, we expand the repertoire of synthetic gene networks built from these regulators by constructing a transcriptional negative autoregulation (NAR) network out of small RNAs (sRNAs). NAR network motifs are core motifs of natural genetic networks, and are known for reducing network response time and steady state signal. Here we use cell-free transcription-translation (TX-TL) reactions and a computational model to design and prototype sRNA NAR constructs. Using parameter sensitivity analysis, we design a simple set of experiments that allow us to accurately predict NAR function in TX-TL. We transfer successful network designs into Escherichia coli and show that our sRNA transcriptional network reduces both network response time and steady-state gene expression. This work broadens our ability to construct increasingly sophisticated RNA genetic networks with predictable function.

Original languageEnglish (US)
Pages (from-to)1507-1518
Number of pages12
JournalACS synthetic biology
Volume7
Issue number6
DOIs
StatePublished - Jun 15 2018

Keywords

  • RNA synthetic circuitry
  • model-guided design
  • negative autoregulation
  • parameterization
  • sensitivity analysis
  • transcriptional sRNA regulator

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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