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Updated: Jun 18, 2026

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Sensitive and Broadly Compatible Transcription Factor-Based Biosensor for Monitoring c-di-GMP Dynamics in Biofilms.

Yidan Hu1,2, Jian Liu1, Aloysius Teng2,3

  • 1Department of Biological Sciences and Technology, School of Environmental Studies, China University of Geosciences, Wuhan 430074, P. R. China.

ACS Synthetic Biology
|May 16, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel biosensor to detect cyclic di-GMP (c-di-GMP) levels, crucial for understanding and mitigating harmful biofilms. This tool aids in developing new strategies to control biofilm formation and growth.

Keywords:
biofilmscyclic di-GMPsecond messengertranscription factor FleQtranscription factor-based biosensor

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Area of Science:

  • Microbiology and Molecular Biology
  • Biotechnology
  • Biofilm Research

Background:

  • Biofilms are widespread microbial communities with detrimental impacts, including infections and biocorrosion.
  • The second messenger cyclic di-GMP (c-di-GMP) plays a pivotal role in regulating biofilm formation.
  • Reducing intracellular c-di-GMP levels is a key strategy for developing effective biofilm mitigation approaches.

Purpose of the Study:

  • To develop a transcription factor (TF)-based biosensor for detecting and quantifying intracellular c-di-GMP levels.
  • To optimize the biosensor for broad compatibility across diverse bacterial species.
  • To utilize the biosensor for monitoring c-di-GMP dynamics in real-time within biofilms.

Main Methods:

  • Development of a TF-based biosensor integrating FleQ TF with a P-P tandem promoter.
  • Optimization of TF expression to fine-tune the biosensor's dynamic range.
  • Ratiometric, image-based quantification using green fluorescence protein (GFP) as a reporter and DAPI or mRFP for biomass, calculating GFP/DAPI or GFP/mRFP ratios.

Main Results:

  • The biosensor demonstrated broad compatibility and effectiveness in detecting reduced c-di-GMP levels in various model organisms, including those lacking FleQ or its homologues.
  • Successful monitoring of c-di-GMP levels in *Pseudomonas aeruginosa* and *Vibrio cholerae* biofilms.
  • The developed methodology provides a measure of effective c-di-GMP per unit of biofilm biomass.

Conclusions:

  • The developed TF-based biosensor is a valuable tool for studying c-di-GMP dynamics in bacteria.
  • This biosensor facilitates research into regulatory networks governing biofilm development.
  • The tool supports the development of novel strategies for controlling problematic biofilms.