Related Experiment Videos
A novel drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR)
P Honkakoski1, I Jääskeläinen, M Kortelahti
1Department of Pharmaceutics, University of Kuopio, Finland. paavo.honkakoski@uku.fi
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.