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Nat Commun. 2020 Jan 30;11(1):605. doi: 10.1038/s41467-020-14460-4.

Optogenetic regulation of endogenous proteins.

Author information

1
Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland.
2
Institute of Biotechnology, HiLIFE, University of Helsinki, Helsinki, 00790, Finland.
3
Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, 10461, USA.
4
Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland. vladislav.verkhusha@einsteinmed.org.
5
Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, 10461, USA. vladislav.verkhusha@einsteinmed.org.

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.

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