Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET

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dc.contributor.author Shcherbakova, Daria M.
dc.contributor.author Cammer, Natasha Cox
dc.contributor.author Huisman, Tsipora M.
dc.contributor.author Verkhusha, Vladislav V.
dc.contributor.author Hodgson, Louis
dc.date.accessioned 2019-07-11T21:36:58Z
dc.date.available 2021-12-17T22:02:50Z
dc.date.issued 2018-06
dc.identifier.citation Shcherbakova , D M , Cammer , N C , Huisman , T M , Verkhusha , V V & Hodgson , L 2018 , ' Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET ' , Nature Chemical Biology , vol. 14 , no. 6 , pp. 591-+ . https://doi.org/10.1038/s41589-018-0044-1
dc.identifier.other PURE: 109386371
dc.identifier.other PURE UUID: c7a9f4b0-7d92-4db2-8d88-0d9ae82c1f8d
dc.identifier.other WOS: 000435445100016
dc.identifier.other Scopus: 85045851039
dc.identifier.uri http://hdl.handle.net/10138/303899
dc.description.abstract Direct visualization and light control of several cellular processes is a challenge, owing to the spectral overlap of available genetically encoded probes. Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Forster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools. We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways. Specifically, we combined the Rac1 biosensor with CFP-YFP FRET biosensors for RhoA and for Rac1-GDI binding, and concurrently used the LOV-TRAP tool for upstream Rac1 activation. We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK; showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules; and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1. en
dc.format.extent 17
dc.language.iso eng
dc.relation.ispartof Nature Chemical Biology
dc.rights unspecified
dc.rights.uri info:eu-repo/semantics/openAccess
dc.subject FLUORESCENT PROTEINS
dc.subject LIVING CELLS
dc.subject IN-VIVO
dc.subject SPATIOTEMPORAL DYNAMICS
dc.subject BACTERIAL PHYTOCHROME
dc.subject CDC42 ACTIVATION
dc.subject BIOSENSORS
dc.subject RAC
dc.subject TOOLS
dc.subject COORDINATION
dc.subject 1182 Biochemistry, cell and molecular biology
dc.subject 3111 Biomedicine
dc.title Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET en
dc.type Article
dc.contributor.organization Medicum
dc.contributor.organization Department of Biochemistry and Developmental Biology
dc.description.reviewstatus Peer reviewed
dc.relation.doi https://doi.org/10.1038/s41589-018-0044-1
dc.relation.issn 1552-4450
dc.rights.accesslevel openAccess
dc.type.version publishedVersion

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