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Temperature-responsive optogenetic probes of cell signaling
Journal article   Open access   Peer reviewed

Temperature-responsive optogenetic probes of cell signaling

William Benman, Erin E. Berlew, Hao Deng, Caitlyn Parker, Ivan A. Kuznetsov, Bomyi Lim, Arndt F. Siekmann, Brian Y. Chow and Lukasz J. Bugaj
Nature chemical biology, v 18(2), pp 152-160
01 Feb 2022
PMID: 34937907
url
https://www.ncbi.nlm.nih.gov/pmc/articles/9252025View
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Abstract

631/136/334/1874/763 631/553 631/80/86 631/92/96 Article Biochemical Engineering Biochemistry Bioorganic Chemistry Cell Biology Chemistry Chemistry and Materials Science Chemistry/Food Science General
We describe single-component optogenetic probes whose activation dynamics depend on both light and temperature. We used the BcLOV4 photoreceptor to stimulate Ras and phosphatidyl inositol-3-kinase signaling in mammalian cells, allowing activation over a large dynamic range with low basal levels. Surprisingly, we found that BcLOV4 membrane translocation dynamics could be tuned by both light and temperature such that membrane localization spontaneously decayed at elevated temperatures despite constant illumination. Quantitative modeling predicted BcLOV4 activation dynamics across a range of light and temperature inputs and thus provides an experimental roadmap for BcLOV4-based probes. BcLOV4 drove strong and stable signal activation in both zebrafish and fly cells, and thermal inactivation provided a means to multiplex distinct blue-light sensitive tools in individual mammalian cells. BcLOV4 is thus a versatile photosensor with unique light and temperature sensitivity that enables straightforward generation of broadly applicable optogenetic tools. The BcLOV4 photoreceptor was used to generate single-component optogenetic signaling probes whose activation dynamics are dependent on light and temperature, allowing multiplexing of blue-light-sensitive tools in mammalian cells

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Biochemistry & Molecular Biology
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