Modulating Optogenetic YAP In Vitro and In Vivo.

Area of Science:

• Cell Biology
• Molecular Biology
• Biotechnology

Background:

• The Hippo-YAP signaling pathway regulates organ growth and regeneration.
• YAP signaling dysregulation contributes to cancer metastasis through uncontrolled proliferation and invasion.
• Precise temporal control of YAP activity is crucial for studying its role in biological processes.

Purpose of the Study:

• To develop an optogenetic system for controlling YAP subcellular localization and activity.
• To enable second-to-minute timescale manipulation of YAP function.
• To investigate YAP's role in biological processes with enhanced temporal precision.

Main Methods:

• Engineered an optogenetic construct using LOV2-Jα interacting domains to photocage a nuclear localization signal (NLS) fused to YAP.
• Utilized 488 nm light to induce Jα helix unfolding, disrupting LOV2 interaction and exposing the NLS.
• Demonstrated light-induced nuclear translocation of the YAP optogenetic construct.

Main Results:

• Successfully achieved light-inducible nuclear shuttling of the YAP construct.
• Confirmed that the nuclear translocation is reversible and tuneable.
• Validated functional YAP activity in vitro and in vivo following nuclear localization.

Conclusions:

• The developed optogenetic system provides precise spatiotemporal control over YAP activity.
• This tool facilitates detailed investigation of the Hippo-YAP pathway in various biological contexts.
• The system holds potential for studying YAP's role in development, regeneration, and cancer.