Abstract
The constitutive androstane receptor (CAR, NR1I3) controls the transcription of numerous hepatic drug metabolizing enzymes and transporters. There are two possible methods of activation for CAR, direct ligand binding and a ligand-independent method, which makes this a unique nuclear receptor. Both mechanisms require the translocation of CAR from the cytoplasm into the nucleus. Interestingly, CAR is constitutively active and spontaneously localized in the nucleus of most immortalized cell lines. This creates an important challenge in most in vitro assay models because immortalized cells cannot be used without inhibiting the high basal activity. In this book chapter, we go into detail of how to perform quantitative high-throughput screens to identify human CAR modulators through the employment of a double stable cell line. Using this line, we can identify activators, as well as deactivators, of the challenging nuclear receptor, CAR.