نبذة مختصرة : Techniques of protein regulation, such as conditional gene expression, RNA interference, knock-in and knock-out, lack sufficient spatiotemporal accuracy, while optogenetic tools suffer from non-physiological response due to overexpression artifacts. Here we present a near-infrared light-activatable optogenetic system, which combines the specificity and orthogonality of intrabodies with the spatiotemporal precision of optogenetics. We engineer optically-controlled intrabodies to regulate genomically expressed protein targets and validate the possibility to further multiplex protein regulation via dual-wavelength optogenetic control. We apply this system to regulate cytoskeletal and enzymatic functions of two non-tagged endogenous proteins, actin and RAS GTPase, involved in complex functional networks sensitive to perturbations. The optogenetically-enhanced intrabodies allow fast and reversible regulation of both proteins, as well as simultaneous monitoring of RAS signaling with visible-light biosensors, enabling all-optical approach. Growing number of intrabodies should make their incorporation into optogenetic tools the versatile technology to regulate endogenous targets. Optogenetic approaches to control protein-protein interactions usually require overexpression of the target proteins. Here the authors integrate intrabodies into near-infrared- and blue-light activatable optogenetic tools to control endogenous proteins in mammalian cells. ; Peer reviewed
Relation: Redchuk , T A , Karasev , M M , Verkhusha , P V , Donnelly , S K , Hülsemann , M , Virtanen , J , Moore , H M , Vartiainen , M K , Hodgson , L & Verkhusha , V V 2020 , ' Optogenetic regulation of endogenous proteins ' , Nature Communications , vol. 11 , no. 1 , 605 . https://doi.org/10.1038/s41467-020-14460-4; Bibtex: redchuk2020optogenetic; ORCID: /0000-0002-2017-0475/work/77847329; ORCID: /0000-0002-3966-7908/work/77848743; ad5fbbc7-7fa5-4af5-8f50-b486ca5d0aa1; http://hdl.handle.net/10138/317811; 000543984200004
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