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Updated: Jan 25, 2026

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
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Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

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A Reverse Transfection Method for Screening of Nuclear Receptor Activators

Jenni Küblbeck1, Paavo Honkakoski2

  • 1Faculty of Health Sciences, School of Pharmacy, University of Eastern Finland, Kuopio, Finland. jenni.kublbeck@uef.fi.

Methods in Molecular Biology (Clifton, N.J.)
|May 2, 2019
PubMed

Insights

This study introduces a reverse transfection system for studying nuclear receptor activation. This method offers stable, uniform, and reproducible results for drug development and gene expression studies.

Area of Science:

  • Pharmacology
  • Molecular Biology
  • Biochemistry

Background:

  • Reporter assays are crucial for studying nuclear receptor activation.
  • Evaluating drug candidates' potential to induce cytochrome P450 enzymes is vital in drug development.
  • Conventional transfection methods can introduce variability in experimental results.

Purpose of the Study:

  • To describe a novel reverse transfection protocol for studying human nuclear receptors.
  • To investigate the activation of constitutive androstane receptor (CAR) and pregnane X receptor (PXR).
  • To provide a reliable method for early drug development and mechanistic studies.

Main Methods:

  • Development of a reverse transfection system.
  • Application of the system to study CAR and PXR activation.
  • Comparison with conventional transfection techniques.

Main Results:

  • The reverse transfection system ensures long-term stability and uniformity of DNA-carrier complexes.
  • This method overcomes the inherent variation seen in conventional transfection approaches.
  • Reproducible and reliable data were obtained for nuclear receptor activation studies.

Conclusions:

  • The described reverse transfection system is a robust tool for studying nuclear receptor activation.
  • It provides a reproducible and adaptable method for early drug development.
  • The system facilitates mechanistic studies on nuclear receptor-mediated gene expression changes.
Keywords:
Constitutive androstane receptorInductionMammalian 1-hybridPregnane X receptorReporter geneReverse transfection

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