Optogenetic control of protein binding using light-switchable nanobodies. Author Agnieszka Gil, César Carrasco-López, Liyuan Zhu, Evan Zhao, Pavithran Ravindran, Maxwell Wilson, Alexander Goglia, José Avalos, Jared Toettcher Publication Year 2020 Type Journal Article Abstract A growing number of optogenetic tools have been developed to reversibly control binding between two engineered protein domains. In contrast, relatively few tools confer light-switchable binding to a generic target protein of interest. Such a capability would offer substantial advantages, enabling photoswitchable binding to endogenous target proteins in cells or light-based protein purification in vitro. Here, we report the development of opto-nanobodies (OptoNBs), a versatile class of chimeric photoswitchable proteins whose binding to proteins of interest can be enhanced or inhibited upon blue light illumination. We find that OptoNBs are suitable for a range of applications including reversibly binding to endogenous intracellular targets, modulating signaling pathway activity, and controlling binding to purified protein targets in vitro. This work represents a step towards programmable photoswitchable regulation of a wide variety of target proteins. Keywords Animals, Mice, Humans, Signal Transduction, Protein Binding, HEK293 Cells, NIH 3T3 Cells, Optogenetics, Protein Transport, Synthetic Biology Journal Nat Commun Volume 11 Issue 1 Pages 4044 Date Published 2020 Aug 13 ISSN Number 2041-1723 DOI 10.1038/s41467-020-17836-8 Alternate Journal Nat Commun PMCID PMC7426870 PMID 32792536 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML