Receptor gene assays

Oestrogen receptor binding/Reporter gene assay. 

Legler J, van den Brink CE, Brouwer A, et al. 1999. Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line. Toxicol Sci 48 55-66. 

The yeast reporter gene assays not only assess for the interaction of the chemical with the hormone receptor, but also the ability of that receptor-chemical ligand interaction to activate the hormone DNA response element. It should be realized, however, that most of these systems have been developed with human and mammalian hormone receptors and differences in ligand potencies can occur between different animal species. A comprehensive review of in vitro assays for measuring estrogenic activity, and some of the issues of comparability, is provided by Zacharewski (1997). 

Assays can be prepared with a reporter system containing, for example, the hrefly luciferase gene. The reporter cells are coupled to receptor genes. When a ligand binds to the receptor, a luminescent glow can be observed. In this way, the effects of the signaling events are evaluated. 

OECD. 2006e. Stably transfected transcriptional activation TA) assay for detecting estrogenic activity of chemicals - The human estrogen receptor alpha mediated reporter gene assay using HeLa-hER-9903 cell line. Version October 2006. Paris OECD, http //www.oecd.org/dataoecd/49/55/37531918.pdf... 

Freitas, J., Cano, P, Craig-Veit, C., Goodson, M.L., Furlowe, Murk, A.J. (2009). Detection of thyroid hormone receptor disrupters by a novel stable in vitro reporter gene assay. Submitted. 

Wnendt, S., Kriiger, T., Janocha, E., Hildebrandt, D, Engiberger, W. Agonistic effect of buprenorphine in a nociceptin/OFQ receptor triggered reporter gene assay, Mol. Pharmacol. 1999, 56, 334-338. 

Gaido, K.W., Leonard, L.S., Lovell, S., Gould, J.C., Babai, D., Portier, C.J., and McDonnell, D.P., Evaluation of chemicals with endocrine modulating activity in a yeast-based steroid hormone receptor gene transcription assay, Toxicol. Appl. Pharmacol., 143, 205-212, 1997. 

Development of Reporter Gene Assay for Calcitonin Receptor.67... 

Calcitonin Receptor (5-Lactamase Reporter Gene Assay... 

Figure 2.9. Luciferase reporter gene assay/promoter deletion analysis. Cells are transfected with the CYP1A1 promoter reporter (luciferase) constructs in which the promoter region is increasing deleted. The cells are then treated with benzo[a]pyrene (an inducer of CYP1A1), and cells are harvested 24-48hr later and luciferase activity is measured. Promoter deletion analysis revealed enhancer element between -1140 and -1029. Contained within this region is xenobiotic response element (XRE) to which the aryl hydrocarbon receptor (AHR) binds.

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