Predictive Spatiotemporal Manipulation of Signaling Perturbations Using Optogenetics |
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| Authors: | Leo Valon Fred Etoc Amanda Remorino Florencia di?Pietro Xavier Morin Maxime Dahan Mathieu Coppey |
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| Affiliation: | 1.Laboratoire Physico-Chimie, Institut Curie, Centre National de la Recherche Scientifique UMR168, Paris-Science Lettres, Université Pierre et Marie Curie-Paris 6, Paris, France;2.Center for Studies in Physics and Biology, The Rockefeller University, New York, New York;3.Institut de Biologie de l''École Normale Supérieure, Ecole Normale Supérieure, Paris, France |
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| Abstract: | Recently developed optogenetic methods promise to revolutionize cell biology by allowing signaling perturbations to be controlled in space and time with light. However, a quantitative analysis of the relationship between a custom-defined illumination pattern and the resulting signaling perturbation is lacking. Here, we characterize the biophysical processes governing the localized recruitment of the Cryptochrome CRY2 to its membrane-anchored CIBN partner. We develop a quantitative framework and present simple procedures that enable predictive manipulation of protein distributions on the plasma membrane with a spatial resolution of 5 μm. We show that protein gradients of desired levels can be established in a few tens of seconds and then steadily maintained. These protein gradients can be entirely relocalized in a few minutes. We apply our approach to the control of the Cdc42 Rho GTPase activity. By inducing strong localized signaling perturbation, we are able to monitor the initiation of cell polarity and migration with a remarkable reproducibility despite cell-to-cell variability. |
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