Enhanced Detection of Estrogen-like Compounds by Genetically Engineered Yeast Sensor Strains.
Nidaa Abu-RmailahLiat MoscoviciCarolin RiegrafHadas AtiasSebastian BuchingerGeorg ReifferscheidShimshon BelkinPublished in: Biosensors (2024)
The release of endocrine-disrupting compounds (EDCs) to the environment poses a health hazard to both humans and wildlife. EDCs can activate or inhibit endogenous endocrine functions by binding hormone receptors, leading to potentially adverse effects. Conventional analytical methods can detect EDCs at a high sensitivity and precision, but are blind to the biological activity of the detected compounds. To overcome this limitation, yeast-based bioassays have previously been developed as a pre-screening method, providing an effect-based overview of hormonal-disruptive activity within the sample prior to the application of analytical methods. These yeast biosensors express human endocrine-specific receptors, co-transfected with the relevant response element fused to the specific fluorescent protein reporter gene. We describe several molecular manipulations of the sensor/reporter circuit in a Saccharomyces cerevisiae bioreporter strain that have yielded an enhanced detection of estrogenic-like compounds. Improved responses were displayed both in liquid culture (96-well plate format) as well as in conjunction with sample separation using high-performance thin-layer chromatography (HPTLC). The latter approach allows for an assessment of the biological effect of individual sample components without the need for their chemical identification at the screening stage.
Keyphrases
- saccharomyces cerevisiae
- liquid chromatography
- label free
- crispr cas
- mass spectrometry
- public health
- healthcare
- endothelial cells
- loop mediated isothermal amplification
- gene expression
- cell wall
- real time pcr
- tandem mass spectrometry
- type diabetes
- copy number
- social media
- climate change
- high performance liquid chromatography
- pluripotent stem cells
- amino acid