Monitoring Hippo Signaling Pathway Activity Using a Luciferase-Based Large Tumor Suppressor (LATS) Biosensor

Abstract

Kinase cascades are a fundamental feature of cellular signaling and play a vital role in disease progression. Thus, tools to monitor the activity of kinase cascades are of high importance. Our group has developed a split-luciferase biosensor system to monitor the activity of the Hippo pathway, a kinase cascade that regulates a wide variety of cellular processes. We have applied this biosensor for high-throughput screening to discover novel upstream kinase regulators of Hippo signaling. In this chapter, we describe our approach to designing, validating, and using the biosensor for screening procedures, which provides an example for the reader should they wish to design a similar biosensor system for their own purposes.