Optogenetic regulation of endogenous proteins

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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

Title: Optogenetic regulation of endogenous proteins
Author: Redchuk, Taras A; Karasev, Maksim M; Verkhusha, Polina V; Donnelly, Sara K; Hülsemann, Maren; Virtanen, Jori; Moore, Henna M; Vartiainen, Maria K; Hodgson, Louis; Verkhusha, Vladislav V
Contributor: University of Helsinki, Institute of Biotechnology
University of Helsinki, Department of Anatomy
University of Helsinki, Machine learning in biomedicine
University of Helsinki, Research Services
University of Helsinki, Institute of Biotechnology
University of Helsinki, Department of Anatomy
Date: 2020-01-30
Language: eng
Number of pages: 12
Belongs to series: Nature Communications
ISSN: 2041-1723
URI: http://hdl.handle.net/10138/317811
Abstract: 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.
Subject: 3111 Biomedicine
1182 Biochemistry, cell and molecular biology
GTPASE ACTIVITIES
CELL
RAS
ANTIBODIES
LOCALIZATION
GROWTH
INACTIVATION
INHIBITION
BIOSENSORS
SEQUENCE
GTPASE ACTIVITIES
CELL
RAS
ANTIBODIES
LOCALIZATION
GROWTH
INACTIVATION
INHIBITION
BIOSENSORS
SEQUENCE
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