A novel drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR)

Pharm Res. 2001 Feb;18(2):146-50. doi: 10.1023/a:1011068015301.

Abstract

Purpose: To develop and characterize a new drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR).

Methods: Both transient and stable transfection into HEK293 cells of luciferase plasmids under the control of either drug- and steroid-responsive nuclear receptor CAR or the tetracycline-sensitive transactivator tTA were used in development of stable cell lines.

Results: A stable first-generation cell line that expresses luciferase gene under the control of nuclear receptor CAR was developed. The luciferase expression in CAR-producing cells could be suppressed by androstanes and reactivated by structurally unrelated drugs chlorpromazine, metyrapone, phenobarbital, and clotrimazole. The kinetics of luciferase expression in CAR-producing cells and the tTA system were comparable. The overall regulation of CAR system was improved by modifications to the DNA binding domain and site.

Conclusions: Because of its wide ligand selectivity and transferable ligand binding domain, CAR expands the repertoire of regulated gene expression systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androstenols / pharmacology
  • Binding Sites
  • Biotechnology / methods*
  • Cells, Cultured
  • Constitutive Androstane Receptor
  • DNA / drug effects
  • DNA / metabolism
  • Gene Expression Regulation* / drug effects
  • Genes, Reporter* / drug effects
  • Humans
  • Ligands
  • Luciferases / genetics
  • Pyridines / pharmacology
  • Receptors, Cytoplasmic and Nuclear / genetics*
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • Androstenols
  • Constitutive Androstane Receptor
  • Ligands
  • Pyridines
  • Receptors, Cytoplasmic and Nuclear
  • Transcription Factors
  • 1,4-bis(2-(3,5-dichloropyridyloxy))benzene
  • DNA
  • Luciferases