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Optogenetics pp 363-376 | Cite as

Protein Inactivation by Optogenetic Trapping in Living Cells

  • Hyerim Park
  • Sangkyu Lee
  • Won Do HeoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1408)

Abstract

Optogenetic modules that use genetically encoded elements to control protein function in response to light allow for precise spatiotemporal modulation of signaling pathways. As one of optical approaches, LARIAT (Light-Activated Reversible Inhibition by Assembled Trap) is a unique light-inducible inhibition system that reversibly sequesters target proteins into clusters, generated by multimeric proteins and a blue light-induced heterodimerization module. Here we present a method based on LARIAT for optical inhibition of targets in living mammalian cells. In the protocol, we focus on the inhibition of proteins that modulate cytoskeleton and cell cycle, and describe how to transfect, conduct a photo-stimulation, and analyze the data.

Key words

LARIAT Protein inhibition Optogenetics Cryptochrome 2 (CRY2) CIB1 Clustering Cytoskeleton Cell division 

Notes

Acknowledgements

This work was supported by the Institute for Basic Science (no. IBS-R001-G1), Republic of Korea. Figures 1, 2, 4, 5, and 8 are reproduced with permission from Nature Methods.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Biological SciencesKorea Advanced Institute of Science and Technology (KAIST)DaejeonRepublic of Korea
  2. 2.Center for Cognition and SocialityInstitute for Basic Science (IBS)DaejeonRepublic of Korea
  3. 3.Center for Cognition and SocialityInstitute for Basic Science (IBS)SeoulRepublic of Korea
  4. 4.Department of Biological SciencesKorea Advanced Institute of Science and TechnologyDaejeonRepublic of Korea

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