Optogenetic regulation of endogenous proteins

Taras A. Redchuk, Maksim M. Karasev, Polina V. Verkhusha, Sara K. Donnelly, Maren Hülsemann, Jori Virtanen, Henna M. Moore, Maria K. Vartiainen, Louis Hodgson and Vladislav V. Verkhusha ()
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Taras A. Redchuk: University of Helsinki
Maksim M. Karasev: University of Helsinki
Polina V. Verkhusha: Albert Einstein College of Medicine
Sara K. Donnelly: Albert Einstein College of Medicine
Maren Hülsemann: Albert Einstein College of Medicine
Jori Virtanen: University of Helsinki
Henna M. Moore: University of Helsinki
Maria K. Vartiainen: University of Helsinki
Louis Hodgson: Albert Einstein College of Medicine
Vladislav V. Verkhusha: University of Helsinki

Nature Communications, 2020, vol. 11, issue 1, 1-12

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-14460-4

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