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

Research output: Contribution to journalArticleScientificpeer-review

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.

Original languageEnglish
Article number605
JournalNature Communications
Volume11
Issue number1
Number of pages12
ISSN2041-1723
DOIs
Publication statusPublished - 30 Jan 2020
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 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

Cite this