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Updated: May 11, 2026
A Fluorescence-Based Assay of Membrane Potential for High-Throughput Functional Study of Two Endogenous Ion Channels in Two Epithelial Cell Lines
Published on: June 22, 2022
A rapid membrane potential assay to monitor CFTR function and inhibition
Rangan Maitra1, Perumal Sivashanmugam, Keith Warner
11RTI International, Research Triangle Park, NC, USA.
Abstract:
The cystic fibrosis transmembrane conductance regulator (CFTR) protein is an important regulator of ion transport and fluid secretion in humans. Mutations to CFTR cause cystic fibrosis, which is a common recessive genetic disorder in Caucasians. Involvement of CFTR has been noted in other important diseases, such as secretory diarrhea and polycystic kidney disease. The assays to monitor CFTR function that have been described to date either are complicated or require specialized instrumentation and training for execution. In this report, we describe a rapid FlexStation-based membrane potential assay to monitor CFTR function. In this assay, agonist-mediated activation of CFTR results in membrane depolarization that can be monitored using a fluorescent membrane potential probe. Availability of a simple mix-and-read assay to monitor the function of this important protein might accelerate the discovery of CFTR ligands to study a variety of conditions.
Insights
A new, simple assay measures cystic fibrosis transmembrane conductance regulator (CFTR) protein function. This rapid method may accelerate the discovery of drugs for cystic fibrosis and related conditions.
Area of Science:
- Biochemistry
- Molecular Biology
- Physiology
Background:
- The cystic fibrosis transmembrane conductance regulator (CFTR) protein is crucial for ion transport and fluid secretion.
- CFTR mutations cause cystic fibrosis, a common genetic disorder, and are implicated in other diseases like secretory diarrhea and polycystic kidney disease.
- Existing assays for CFTR function are often complex and require specialized equipment and training.
Purpose of the Study:
- To develop a rapid and accessible assay for monitoring CFTR protein function.
- To facilitate the discovery of novel CFTR ligands for therapeutic research.
Main Methods:
- A FlexStation-based membrane potential assay was developed.
- The assay utilizes a fluorescent membrane potential probe to detect agonist-mediated CFTR activation.
- CFTR activation leads to membrane depolarization, which is quantifiable.
Main Results:
- The described assay provides a simple, mix-and-read method for assessing CFTR function.
- This assay simplifies the monitoring of CFTR activity compared to existing methods.
- The assay's ease of use may expedite the identification of compounds affecting CFTR.
Conclusions:
- A novel, rapid FlexStation-based membrane potential assay for CFTR function has been established.
- This assay offers a simplified approach to studying CFTR, potentially accelerating drug discovery.
- The developed assay could be instrumental in advancing research for cystic fibrosis and related disorders.
