The innate immune response is mediated by cellular recognition of invading pathogens <i>via</i> Pattern Recognition Receptors (PRRs). Although spatial and temporal elements of activation mediate the intensity of the immune response, few tools exist to directly examine these effects. Optogenetics is a convenient methodology for restricting protein activation with spatiotemporal precision. To elucidate the spatiotemporal aspects of innate immune responses, we have designed an optogenetic pattern recognition receptor which activates in response to blue light. We demonstrate direct receptor activation leading to spatial and temporal control of downstream signaling pathways in a variety of relevant cell types. Augmenting our platform with Bi-molecular Fluorescence Complementation (BiFC) results in selective labeling of cells in which receptor activation has occurred, enabling analysis of immune signaling on the single-cell level. In addition, genetic integration enables the examination of receptor activation in a specific cell or tissue type. This information will guide adjuvant design by elucidating the mechanism of single-cell immune activation and enabling a greater understanding of how specific cells communicate pathogen invasion.<br/>

Photoactivatable innate immune receptor for optogenetic inflammation

253rd National Meeting (2017)

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