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National Institute of Environmental Health Sciences

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Your Environment. Your Health.

Publication Detail

Title: Third-generation Ah receptor-responsive luciferase reporter plasmids: amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness.

Authors: He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S

Published In Toxicol Sci, (2011 Oct)

Abstract: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene-based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts.

PubMed ID: 21775728 Exiting the NIEHS site

MeSH Terms: Animals; Biological Assay; Carcinoma, Hepatocellular/drug therapy*; Carcinoma, Hepatocellular/metabolism; Cell Line, Tumor; Clone Cells; Environmental Monitoring/methods; Enzyme Induction; Humans; Luciferases/biosynthesis; Luciferases/genetics*; Mice; Plasmids/genetics*; Predictive Value of Tests; Rats; Receptors, Aryl Hydrocarbon/drug effects; Receptors, Aryl Hydrocarbon/genetics*; Receptors, Aryl Hydrocarbon/metabolism; Reproducibility of Results; Response Elements/drug effects; Response Elements/genetics*; Species Specificity; Tetrachlorodibenzodioxin/pharmacology*; Transcription, Genetic/drug effects; Transfection; Water Pollutants, Chemical/analysis; Water Pollutants, Chemical/toxicity

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