Related Experiment Video
Updated: May 12, 2025
09:07
Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
Published on: December 19, 2018
6.3K
Rewiring Estrogen Receptor α into Bisphenol Selective Receptors Using Darwin Assembly-Based Directed Evolution (DADE)
Roy Eerlings1,2,3, Xiao Yin Lee1, Wout Van Eynde4
1Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
ACS Synthetic Biology
|May 10, 2025
Summary
Researchers engineered yeast estrogen receptors to create sensitive bioassays for detecting bisphenols. This advancement aids in monitoring environmental contaminants and protecting human health and ecosystems.
Area of Science:
- Biotechnology
- Environmental Science
- Molecular Biology
Background:
- Bisphenols are prevalent in plastics, posing environmental and health risks.
- Existing detection methods are limited in scope and sensitivity.
- Current bioassays for estrogenic activity lack specificity and are prone to background interference.
Purpose of the Study:
- To develop a highly sensitive and specific yeast-based bioassay for bisphenols.
- To engineer estrogen receptor variants for enhanced bisphenol recognition.
Main Methods:
- Directed evolution and multi-site mutagenesis of estrogen receptor alpha in Saccharomyces cerevisiae.
- Creation and characterization of preBASE and BASE variants.
- Ligand binding assays with purified ligand-binding domains.
Main Results:
- Identified the preBASE variant with increased bisphenol A sensitivity and reduced estrogen response.
- Developed the BASE variant, a novel bisphenol receptor with primary agonist activity for bisphenols.
- Confirmed receptor rewiring through ligand binding assays.
Conclusions:
- Engineered yeast estrogen receptors serve as a foundation for developing specific, high-throughput bisphenol detection assays.
- This work provides crucial tools for environmental monitoring and risk assessment of bisphenol exposure.
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