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Updated: Mar 29, 2026

Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter
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Identification of Transcription Factor Regulators using Medium-Throughput Screening of Arrayed Libraries and a Dual-Luciferase-Based Reporter

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Activation of Yap-Directed Transcription by Knockdown of Conserved Cellular Functions

C Agarinis1, V Orsini1, P Megel1

  • 1Novartis Institutes for Biomedical Research, Basel, Switzerland.

Journal of Biomolecular Screening
|December 6, 2015
PubMed

Insights

Researchers identified new regulators of the Yap-Hippo pathway using a siRNA screen. This pathway controls cell growth and organ size, and understanding its upstream regulators is key for regeneration research.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The Yap-Hippo pathway is crucial for regulating cell proliferation, growth, and organ size.
  • The pathway involves transcription coactivators YAP and TAZ, but its upstream regulators remain largely uncharacterized.

Purpose of the Study:

  • To identify novel regulators of the Yap-Hippo pathway using a siRNA screen in a liver biliary cell line.
  • To discover factors that mediate YAP transcription coactivator activation at high cell density.

Main Methods:

  • A high-content, image-based assay was employed to monitor the intracellular localization of native YAP protein.
  • A siRNA screen was conducted to identify regulators affecting YAP localization.
  • Validation involved quantifying CYR61 mRNA levels and testing compounds targeting identified gene targets.

Main Results:

  • Several validated hits were identified, revealing insights into Yap-Hippo pathway biology.
  • Identified regulators include components of the nuclear pore, influencing YAP cytoplasmic-nuclear shuttling.
  • Proteasomal degradation mechanisms affecting intracellular YAP concentration and localization were also highlighted.

Conclusions:

  • The study successfully identified novel regulators of the Yap-Hippo pathway.
  • Findings elucidate mechanisms of YAP regulation, including nuclear transport and degradation.
  • Targeting conserved cellular functions offers a viable strategy for discovering pathway modulators.
Keywords:
HippoYAPfrequent hitterssiRNA screening

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