Optogenetic control of YAP cellular localisation and function

Abstract

YAP, an effector of the Hippo signalling pathway, promotes organ growth and regeneration. Prolonged YAP activation results in uncontrolled proliferation and cancer. Therefore, exogenous regulation of YAP activity has potential translational applications. We present a versatile optogenetic construct (optoYAP) for manipulating YAP localisation, and consequently its activity and function. We attach a LOV2 domain that photocages a nuclear localisation signal (NLS) to the N‐terminus of YAP. In 488 nm light, the LOV2 domain unfolds, exposing the NLS, which shuttles optoYAP into the nucleus. Nuclear import of optoYAP is reversible and tuneable by light intensity. In cell culture, activated optoYAP promotes YAP target gene expression and cell proliferation. Similarly, optofYap can be used in zebrafish embryos to modulate target genes. We demonstrate that optoYAP can override a cell's response to substrate stiffness to generate anchorage‐independent growth. OptoYAP is functional in both cell culture and in vivo , providing a powerful tool to address basic research questions and therapeutic applications in regeneration and disease. image optoYAP is a new optogenetic tool to manipulate the cellular localisation of YAP and YAP‐dependent gene expression. The efficacy and functionality of this tool is demonstrated in cell culture – including on different substrate stiffnesses – and zebrafish. optoYAP combines both NLS and NES components to ensure rapid nuclear import of YAP upon light‐activation and export of YAP from the nucleus when light is deactivated. optoYAP shows rapid activation and reversal, making it suitable for a range of applications. optoYAP can drive downstream Hippo‐YAP pathway components and further can stimulate growth of non‐transformed cells on soft substrates.