Third-generation ah receptor-responsive luciferase reporter plasmids

Amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness

Guochun He, Tomoaki Tsutsumi, Bin Zhao, David S. Baston, Jing Zhao, Sharon Heath-pagliuso, Michael S. Denison

Research output: Contribution to journalArticle

43 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)511-522
Number of pages12
JournalToxicological Sciences
Volume123
Issue number2
DOIs
StatePublished - Oct 2011

Fingerprint

Dioxins
Bioassay
Luciferases
Biological Assay
Amplification
Plasmids
Screening
Cells
Cell Line
Poisons
Reporter Genes
Gene expression
Transfection
Limit of Detection
Hepatocellular Carcinoma
Genes
Impurities
Gene Expression

Keywords

  • Ah receptor
  • Bioassay
  • CALUX
  • Dioxin
  • TCDD

ASJC Scopus subject areas

  • Toxicology

Cite this

Third-generation ah receptor-responsive luciferase reporter plasmids : Amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness. / He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S.; Zhao, Jing; Heath-pagliuso, Sharon; Denison, Michael S.

In: Toxicological Sciences, Vol. 123, No. 2, 10.2011, p. 511-522.

Research output: Contribution to journalArticle

He, Guochun ; Tsutsumi, Tomoaki ; Zhao, Bin ; Baston, David S. ; Zhao, Jing ; Heath-pagliuso, Sharon ; Denison, Michael S. / Third-generation ah receptor-responsive luciferase reporter plasmids : Amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness. In: Toxicological Sciences. 2011 ; Vol. 123, No. 2. pp. 511-522.
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