TY - JOUR
T1 - A general approach for engineering RTKs optically controlled with far-red light
AU - Leopold, Anna V.
AU - Thankachan, Stephen
AU - Yang, Chun
AU - Gerashchenko, Dmitry
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank J. Ihalainen (University of Jyväskylä, Finland) for the DrBphP gene and D. Lindholm (University of Helsinki, Finland) for the cell lines. This work was supported by the grants GM122567, AG061774 and NS106406 from the US National Institutes of Health and 322226 from the Academy of Finland.
Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2022/7
Y1 - 2022/7
N2 - Regulation of receptor tyrosine kinase (RTK) activity is necessary for studying cell signaling pathways in health and disease. We developed a generalized approach for engineering RTKs optically controlled with far-red light. We targeted the bacterial phytochrome DrBphP to the cell surface and allowed its light-induced conformational changes to be transmitted across the plasma membrane via transmembrane helices to intracellular RTK domains. Systematic optimization of these constructs has resulted in optically regulated epidermal growth factor receptor, HER2, TrkA, TrkB, FGFR1, IR1, cKIT and cMet, named eDrRTKs. eDrRTKs induced downstream signaling in mammalian cells in tens of seconds. The ability to activate eDrRTKs with far-red light enabled spectral multiplexing with fluorescent probes operating in a shorter spectral range, allowing for all-optical assays. We validated eDrTrkB performance in mice and found that minimally invasive stimulation in the neocortex with penetrating via skull far-red light-induced neural activity, early immediate gene expression and affected sleep patterns.
AB - Regulation of receptor tyrosine kinase (RTK) activity is necessary for studying cell signaling pathways in health and disease. We developed a generalized approach for engineering RTKs optically controlled with far-red light. We targeted the bacterial phytochrome DrBphP to the cell surface and allowed its light-induced conformational changes to be transmitted across the plasma membrane via transmembrane helices to intracellular RTK domains. Systematic optimization of these constructs has resulted in optically regulated epidermal growth factor receptor, HER2, TrkA, TrkB, FGFR1, IR1, cKIT and cMet, named eDrRTKs. eDrRTKs induced downstream signaling in mammalian cells in tens of seconds. The ability to activate eDrRTKs with far-red light enabled spectral multiplexing with fluorescent probes operating in a shorter spectral range, allowing for all-optical assays. We validated eDrTrkB performance in mice and found that minimally invasive stimulation in the neocortex with penetrating via skull far-red light-induced neural activity, early immediate gene expression and affected sleep patterns.
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U2 - 10.1038/s41592-022-01517-z
DO - 10.1038/s41592-022-01517-z
M3 - Article
C2 - 35681062
AN - SCOPUS:85131551841
SN - 1548-7091
VL - 19
SP - 871
EP - 880
JO - Nature Methods
JF - Nature Methods
IS - 7
ER -