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Reporter Genes

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Detecting Estrogenic Ligands in Personal Care Products using a Yeast Estrogen Screen Optimized for the Undergraduate Teaching Laboratory

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Published on: January 1, 2018

A new fluorescence complementation biosensor for detection of estrogenic compounds.

Michael J McLachlan¹, John A Katzenellenbogen, Huimin Zhao

¹Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.

Biotechnology and Bioengineering

|July 7, 2011

Summary

Scientists developed a novel fluorescent biosensor to detect environmental estrogenic compounds. This rapid, substrate-free sensor distinguishes between agonists and antagonists, aiding in monitoring endocrine disruptors.

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Area of Science:

Environmental Science
Biotechnology
Molecular Biology

Background:

Estrogenic compounds are prevalent environmental contaminants from diverse sources like pharmaceuticals and waste.
Monitoring these endocrine-disrupting compounds is crucial for environmental and public health.
Existing detection methods may lack speed, specificity, or require substrates.

Purpose of the Study:

To create and validate a novel biosensor for detecting estrogenic compounds.
To develop a system capable of distinguishing between estrogen receptor agonists and antagonists.
To establish a rapid, high-throughput assay for environmental monitoring.

Main Methods:

Engineered split fluorescent protein (mVenus) fragments fused to the estrogen receptor alpha (ERα) ligand-binding domain.
Expressed engineered sensors in HeLa cells for functional testing.
Assessed sensor response (fluorescence) to various estrogenic compounds, including agonists and antagonists.

Main Results:

Sensor 6 (ERα 312-595) demonstrated a nine-fold fluorescence increase with ERα agonists/antagonists.
Sensor 2 (ERα 281-549) differentiated agonists (decreased fluorescence) from antagonists (induced fluorescence).
Sensor 6 showed rapid signal development (2-fold at 4h, 4-fold at 8h) and correlated well with known binding affinities.

Conclusions:

Developed a versatile split fluorescent protein biosensor for estrogenic compounds.
The sensor system is rapid, substrate-free, and can distinguish agonists from antagonists.
This technology offers a promising tool for high-throughput screening of environmental endocrine disruptors.