Establishment of a fluorescence resonance energy transfer-based bioassay for detecting dioxin-like compounds.
Dioxins comprise a group of compounds which contain a double aromatic ring-like structure. They are among the most prevalent and toxic environmental pollutants. Accumulation of dioxins in human tissues poses a potential threat to human health. Currently, analytical chemical procedures dominate dioxin-detection protocols. In this study, we established a fluorescence resonance energy transfer (FRET)-based dioxin-detection bioassay. Aryl hydrocarbon receptor (AHR) and AHR nuclear translocator (ARNT) fused-cyan fluorescent protein (CFP) and -yellow fluorescent protein (YFP) constructed were transiently co-transfected into rat hepatoma cell line, H4IIEC3 cells. Our results showed that no FRET signals were detected in AHR-CFP- and ARNT-YFP-transfected H4IIEC3 cells. However, dioxin treatments upregulated FRET signals in these transfected cells in a dose-dependent manner. This work highlighted the potential of FRET technique in the detection of dioxin-like compounds.
Lin
Chi-Iou
CI
Institute of Zoology, National Taiwan University, No. 1, Roosevelt Rd., Sec. 4, Taipei, 106, Taiwan, ROC.
Hsieh
Chun-Hung
CH
Lee
Stanley Shiao-Ying
SS
Lee
Wei-Shan
WS
Chang-Chien
Gou-Ping
GP
Pan
Chien-Yuan
CY
Lee
Hsinyu
H
D000818:Animals; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D004147:Dioxins / Q000032:analysis*; D004305:Dose-Response Relationship, Drug; D005452:Fluoresceins / Q000032:analysis; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D000072317:Polychlorinated Dibenzodioxins / Q000032:analysis / Q000633:toxicity; D051381:Rats; D018336:Receptors, Aryl Hydrocarbon / Q000235:genetics / Q000378:metabolism; D013723:Teratogens / Q000032:analysis / Q000633:toxicity
D004147:Dioxins; D005452:Fluoresceins; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D013723:Teratogens; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator