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Updated: May 8, 2025
In vivo Optogenetic Stimulation of the Rodent Central Nervous System
Published on: January 15, 2015
Modulating Optogenetic YAP In Vitro and In Vivo
1Mechanobiology Institute, National University of Singapore, Singapore, Singapore. pearlyn.toh@u.nus.edu.
Abstract:
YAP is a central regulator of the Hippo-YAP signaling axis, an evolutionarily conserved pathway that modulates organ growth and regeneration. Dysregulation of YAP signaling leads to uncontrolled proliferation, promoting epithelial-to-mesenchymal transition and invasion in cancer metastasis. Exogenous manipulation of YAP activity at the second-to-minute timescale is an important step in studying the signaling pathway. We present an optogenetic system to control the subcellular localization of YAP and therefore its activity as a transcriptional co-activator. We used the LOV2-Jα interacting domains to photocage a nuclear localization signal (NLS) attached to YAP. Under 488 nm light, the Jα helix unfolds and the interaction with LOV2 is disrupted, thereby exposing the NLS and allowing for the entire optogenetic construct to be shuttled into the nucleus. This nuclear translocation is reversible and tuneable and demonstrates functional activity after nuclear localization both in vitro and in vivo.
Insights
Scientists developed an optogenetic system to control YAP protein activity. This method precisely manipulates YAP
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.
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