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Optogenetic control of YAP cellular localisation and function
Author(s) -
Toh Pearlyn J Y,
Lai Jason K H,
Hermann Anke,
Destaing Olivier,
Sheetz Michael P,
Sudol Marius,
Saunders Timothy E
Publication year - 2022
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.15252/embr.202154401
Subject(s) - optogenetics , microbiology and biotechnology , effector , hippo signaling pathway , cell growth , zebrafish , biology , cell , nuclear localization sequence , hek 293 cells , nucleus , cell culture , gene , neuroscience , genetics
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.