# Enabling High-Throughput Analysis and Single-Cell Imaging of Bacterial Signals

> **NIH NIH R01** · UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH · 2024 · $355,728

## Abstract

PROJECT SUMMARY
 Enabling High-Throughput Analysis and Single-Cell Imaging of Bacterial Signals
Our research aims to understand how bacteria perceive chemical signals to regulate different behaviors. We
have invented different types of biosensors to rapidly measure key signaling molecules in bacteria, including one
for cyclic di-GMP. This signal controls whether bacteria attach to surfaces, form sticky biofilms, and secrete
toxins. One of our major goals is to identify nutrients, other chemicals, and environmental inputs that change
cyclic di-GMP levels in different bacteria. We recently demonstrated a successful approach that combines
structure-based bioinformatics analysis and experimental screening. However, the discovery of primary inputs
remains challenging because each bacterium harbors many cyclic di-GMP signaling enzymes, the signal is
transiently produced, highly charged, and low in abundance, and the screening method remains a key bottleneck.
Thus, this proposal will develop next-generation fluorescent biosensors to enhance high-throughput, quantitative
screening of enzyme activity directly in cells (Aim 2). These biosensors then will be applied to discover primary
inputs for a widespread small molecule binding domain associated with cyclic di-GMP and other signaling
enzymes (Aim 3). In addition, towards understanding environmental factors that regulate cyclic di-GMP, this
proposal will develop a new type of biosensor to perform in situ imaging of cyclic di-GMP in biofilms (Aim 3). In
the long term, this project aims to inform personalized diets to treat inflammatory bowel diseases and promote
gut health.
For this renewal of the project, the original scope also has been expanded to study the permeability of small
molecules into bacterial cells. The permeability process includes passive permeation, active uptake, and active
efflux mechanisms, and is critical to bacterial growth, signaling, and antibiotic resistance. This proposal will
develop a high-throughput assay that enables real-time monitoring of small molecule permeability in cells (Aim
1). The assay will be applied to understand both the molecular structures and genetic factors that affect
accumulation of fluorescent dyes and of clinical antibiotics inside cells. In the long term, this new aim will improve
chemical biology tools that use these dyes and antibiotics treatments.

## Key facts

- **NIH application ID:** 10913403
- **Project number:** 5R01GM124589-08
- **Recipient organization:** UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH
- **Principal Investigator:** Ming Chen Hammond
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $355,728
- **Award type:** 5
- **Project period:** 2017-09-01 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10913403

## Citation

> US National Institutes of Health, RePORTER application 10913403, Enabling High-Throughput Analysis and Single-Cell Imaging of Bacterial Signals (5R01GM124589-08). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10913403. Licensed CC0.

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