Protocol to identify and validate inhibitors of TAZ-TEAD interaction using NanoBiT and TR-FRET biosensors in mammalian cells

TAZ drives breast cancer progression, yet inhibitors targeting its activity remain limited. Here, we present a protocol to identify existing drugs that disrupt TAZ function using a dual NanoLuc Binary Technology (NanoBiT) and time-resolved fluorescence resonance energy transfer (TR-FRET) biosensor. We describe steps for constructing and expressing the biosensor in mammalian cells, optimizing the TR-FRET antibody pair, screening a bioactive compound library in ultra-high throughput format, and validating the drug candidates by NanoBiT and co-immunoprecipitation assays. This biosensor enables the discovery of TAZ-specific drug inhibitors with increased efficiency. For complete details on the use and execution of this protocol, please refer to Ge et al. 1