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, Vladislav V Verkhusha

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review


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.

TidskriftNature Communications
Antal sidor12
StatusPublicerad - 30 jan. 2020
MoE-publikationstypA1 Tidskriftsartikel-refererad


  • 3111 Biomedicinska vetenskaper
  • 1182 Biokemi, cell- och molekylärbiologi

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