A Mechanogenetic Toolkit for Interrogating Cell Signaling in Space and Time

Tools capable of imaging and perturbing mechanical signaling pathways with fine spatiotemporal resolution have been elusive, despite their importance in diverse cellular processes. The challenge in developing a mechanogenetic toolkit (i.e., selective and quantitative activation of genetically encoded mechanoreceptors) stems from the fact that many mechanically activated processes are localized in space and time yet additionally require mechanical loading to become activated. To address this challenge, we synthesized magnetoplasmonic nanoparticles that can image, localize, and mechanically load targeted proteins with high spatiotemporal resolution. We demonstrate their utility by investigating the cell-surface activation of two mechanoreceptors: Notch and vascular endothelial cadherin (VE-cadherin). By measuring cellular responses to a spectrum of spatial, chemical, temporal, and mechanical inputs at the single-molecule and single-cell levels, we reveal how spatial segregation and mechanical force cooperate to direct receptor activation dynamics. This generalizable technique can be used to control and understand diverse mechanosensitive processes in cell signaling. [Display omitted] [Display omitted] •Development of a mechanogenetic single-cell perturbation approach•Interrogation of the spatial, chemical, and mechanical responses of Notch receptors•Identification of the roles of spatial and mechanical cues on VE-cadherin signaling•Spatiotemporal and quantitative control of single-cell transcription by nanoprobes Mechanogenetics, a new approach that uses nanoparticles with imaging, localizing, and mechanically loading capabilities to activate targeted proteins with high spatiotemporal resolution, reveals how chemical, spatial, and mechanical cues cooperate to direct activation dynamics of Notch and VE-cadherin receptors.