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Quantitative High-Throughput Luciferase Screening in Identifying CAR Modulators

  • Caitlin Lynch
  • Jinghua Zhao
  • Hongbing Wang
  • Menghang XiaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1473)

Abstract

The constitutive androstane receptor (CAR, NR1I3) is responsible for the transcription of multiple 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 of these mechanisms require translocation of CAR from the cytoplasm into the nucleus. Interestingly, CAR is constitutively active in immortalized cell lines due to the basal nuclear location of this receptor. 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 hCAR1 modulators through the employment of a double stable cell line. Using this line, we are able to identify activators, as well as deactivators, of the challenging nuclear receptor, CAR.

Key words

Constitutive Androstane Receptor (CAR) Cytochrome P450 2B6 (CYP2B6) Luciferase Quantitative high-throughput screening (qHTS) 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Caitlin Lynch
    • 1
  • Jinghua Zhao
    • 1
  • Hongbing Wang
    • 2
  • Menghang Xia
    • 1
    Email author
  1. 1.National Center for Advancing Translational SciencesNational Institutes of HealthBethesdaUSA
  2. 2.Department of Pharmaceutical SciencesUniversity of Maryland School of PharmacyBaltimoreUSA

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