Engineered orthogonal quorum sensing systems for synthetic gene regulation in escherichia coli

Stefan J. Tekel, Christina L. Smith, Brianna Lopez, Amber Mani, Christopher Connot, Xylaan Livingstone, Karmella Haynes

Research output: Contribution to journalArticle

Abstract

Gene regulators that are controlled by membrane-permeable compounds called homoserine lactones (HSLs) have become popular tools for building synthetic gene networks that coordinate behaviors across populations of engineered bacteria. Synthetic HSL-signaling systems are derived from natural DNA and protein elements from microbial quorum signaling pathways. Crosstalk, where a single HSL can activate multiple regulators, can lead to faults in networks composed of parallel signaling pathways. Here, we report an investigation of quorum sensing components to identify synthetic pathways that exhibit little to no crosstalk in liquid and solid cultures. In previous work, we characterized the response of a single regulator (LuxR) to 10 distinct HSL-synthase enzymes. Our current study determined the responses of five different regulators (LuxR, LasR, TraR, BjaR, and AubR) to the same set of synthases. We identified two sets of orthogonal synthase-regulator pairs (BjaI/BjaR + EsaI/TraR and LasI/LasR + EsaI/TraR) that show little to no crosstalk when they are expressed in Escherichia coli BL21. These results expand the toolbox of characterized components for engineering microbial communities.

Original languageEnglish (US)
Article number80
JournalFrontiers in Bioengineering and Biotechnology
Volume7
Issue numberMAR
DOIs
StatePublished - Jan 1 2019

Fingerprint

Synthetic Genes
Quorum Sensing
Crosstalk
Gene expression
Escherichia coli
Genes
Gene Regulatory Networks
Bacteria
DNA
Regulator Genes
Enzymes
Proteins
Membranes
Liquids
homoserine lactone
Population

Keywords

  • Biosensor
  • Gene circuit
  • Homoserine lactone
  • Quorum sensing
  • Transcription factor

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Histology
  • Biomedical Engineering

Cite this

Engineered orthogonal quorum sensing systems for synthetic gene regulation in escherichia coli. / Tekel, Stefan J.; Smith, Christina L.; Lopez, Brianna; Mani, Amber; Connot, Christopher; Livingstone, Xylaan; Haynes, Karmella.

In: Frontiers in Bioengineering and Biotechnology, Vol. 7, No. MAR, 80, 01.01.2019.

Research output: Contribution to journalArticle

Tekel, Stefan J. ; Smith, Christina L. ; Lopez, Brianna ; Mani, Amber ; Connot, Christopher ; Livingstone, Xylaan ; Haynes, Karmella. / Engineered orthogonal quorum sensing systems for synthetic gene regulation in escherichia coli. In: Frontiers in Bioengineering and Biotechnology. 2019 ; Vol. 7, No. MAR.
@article{383174940e6b48d28773e730163dc050,
title = "Engineered orthogonal quorum sensing systems for synthetic gene regulation in escherichia coli",
abstract = "Gene regulators that are controlled by membrane-permeable compounds called homoserine lactones (HSLs) have become popular tools for building synthetic gene networks that coordinate behaviors across populations of engineered bacteria. Synthetic HSL-signaling systems are derived from natural DNA and protein elements from microbial quorum signaling pathways. Crosstalk, where a single HSL can activate multiple regulators, can lead to faults in networks composed of parallel signaling pathways. Here, we report an investigation of quorum sensing components to identify synthetic pathways that exhibit little to no crosstalk in liquid and solid cultures. In previous work, we characterized the response of a single regulator (LuxR) to 10 distinct HSL-synthase enzymes. Our current study determined the responses of five different regulators (LuxR, LasR, TraR, BjaR, and AubR) to the same set of synthases. We identified two sets of orthogonal synthase-regulator pairs (BjaI/BjaR + EsaI/TraR and LasI/LasR + EsaI/TraR) that show little to no crosstalk when they are expressed in Escherichia coli BL21. These results expand the toolbox of characterized components for engineering microbial communities.",
keywords = "Biosensor, Gene circuit, Homoserine lactone, Quorum sensing, Transcription factor",
author = "Tekel, {Stefan J.} and Smith, {Christina L.} and Brianna Lopez and Amber Mani and Christopher Connot and Xylaan Livingstone and Karmella Haynes",
year = "2019",
month = "1",
day = "1",
doi = "10.3389/fbioe.2019.00080",
language = "English (US)",
volume = "7",
journal = "Frontiers in Bioengineering and Biotechnology",
issn = "2296-4185",
publisher = "Frontiers Media S. A.",
number = "MAR",

}

TY - JOUR

T1 - Engineered orthogonal quorum sensing systems for synthetic gene regulation in escherichia coli

AU - Tekel, Stefan J.

AU - Smith, Christina L.

AU - Lopez, Brianna

AU - Mani, Amber

AU - Connot, Christopher

AU - Livingstone, Xylaan

AU - Haynes, Karmella

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Gene regulators that are controlled by membrane-permeable compounds called homoserine lactones (HSLs) have become popular tools for building synthetic gene networks that coordinate behaviors across populations of engineered bacteria. Synthetic HSL-signaling systems are derived from natural DNA and protein elements from microbial quorum signaling pathways. Crosstalk, where a single HSL can activate multiple regulators, can lead to faults in networks composed of parallel signaling pathways. Here, we report an investigation of quorum sensing components to identify synthetic pathways that exhibit little to no crosstalk in liquid and solid cultures. In previous work, we characterized the response of a single regulator (LuxR) to 10 distinct HSL-synthase enzymes. Our current study determined the responses of five different regulators (LuxR, LasR, TraR, BjaR, and AubR) to the same set of synthases. We identified two sets of orthogonal synthase-regulator pairs (BjaI/BjaR + EsaI/TraR and LasI/LasR + EsaI/TraR) that show little to no crosstalk when they are expressed in Escherichia coli BL21. These results expand the toolbox of characterized components for engineering microbial communities.

AB - Gene regulators that are controlled by membrane-permeable compounds called homoserine lactones (HSLs) have become popular tools for building synthetic gene networks that coordinate behaviors across populations of engineered bacteria. Synthetic HSL-signaling systems are derived from natural DNA and protein elements from microbial quorum signaling pathways. Crosstalk, where a single HSL can activate multiple regulators, can lead to faults in networks composed of parallel signaling pathways. Here, we report an investigation of quorum sensing components to identify synthetic pathways that exhibit little to no crosstalk in liquid and solid cultures. In previous work, we characterized the response of a single regulator (LuxR) to 10 distinct HSL-synthase enzymes. Our current study determined the responses of five different regulators (LuxR, LasR, TraR, BjaR, and AubR) to the same set of synthases. We identified two sets of orthogonal synthase-regulator pairs (BjaI/BjaR + EsaI/TraR and LasI/LasR + EsaI/TraR) that show little to no crosstalk when they are expressed in Escherichia coli BL21. These results expand the toolbox of characterized components for engineering microbial communities.

KW - Biosensor

KW - Gene circuit

KW - Homoserine lactone

KW - Quorum sensing

KW - Transcription factor

UR - http://www.scopus.com/inward/record.url?scp=85065286904&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85065286904&partnerID=8YFLogxK

U2 - 10.3389/fbioe.2019.00080

DO - 10.3389/fbioe.2019.00080

M3 - Article

VL - 7

JO - Frontiers in Bioengineering and Biotechnology

JF - Frontiers in Bioengineering and Biotechnology

SN - 2296-4185

IS - MAR

M1 - 80

ER -