Chapter title |
Protein Inactivation by Optogenetic Trapping in Living Cells
|
---|---|
Chapter number | 25 |
Book title |
Optogenetics
|
Published in |
Methods in molecular biology, March 2016
|
DOI | 10.1007/978-1-4939-3512-3_25 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3510-9, 978-1-4939-3512-3
|
Authors |
Hyerim Park, Sangkyu Lee, Won Do Heo |
Editors |
Arash Kianianmomeni |
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. |
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