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 organization: | Institute of Biotechnology Helsinki Institute of Life Science HiLIFE Medicum Faculty of Medicine University of Helsinki Department of Anatomy Machine learning in biomedicine Research Services Nuclear organization by actin |
Date: | 2020-01-30 |
Language: | eng |
Number of pages: | 12 |
Belongs to series: | Nature Communications |
ISSN: | 2041-1723 |
DOI: | https://doi.org/10.1038/s41467-020-14460-4 |
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 |
Peer reviewed: | Yes |
Rights: | cc_by |
Usage restriction: | openAccess |
Self-archived version: | publishedVersion |
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