Project summary/Abstract Neuroinflammation plays a critical role in neurodegenerative diseases and cognitive functions. Current methods to study neuroinflammation in animal models are extremely limited, primarily restricted to endpoint histological analysis. Furthermore, a means to precisely control the neuroinflammatory response is lacking. The goal of this proposal is to develop the first family of biosensors that can be used to report and dynamically regulate changes in inflammation within the brains of live animals. This is accomplished by a synthetic gene circuit where a sensor of dynamic changes in inflammatory cytokines translates these changes into production of proteins capable of mitigating pathological inflammation. Building on our experience with bioluminescent optogenetics tools, we will develop a light emitting Bioluminescent Kinase Sensor (BlinKS) that activates a light sensing transcription factor driving cytokines to reduce inflammation. Our first aim is to develop and optimize sensors based on split luciferases that will produce light in the presence of a molecule produced in the inflammatory signaling cascade. Inflammation dependent light production is then leveraged to activate light sensitive optogenetic proteins to control cellular activity. Our second aim is to utilize these light emitting sensors of the inflammatory cascade in a genetic circuit to control the expression of an anti-inflammatory cytokine to control inflammation as a self-regulating therapeutic in a transgenic mouse model of neuroinflammation. Our end goal is to apply these inflammation modulators to various animal models where neuroinflammation is a key pathologic hallmark such as Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease. We expect these modulators to have high translational potential to combat neurodegeneration and other inflammatory disorders. This project is enabled by a multidisciplinary team of five labs with expertise in high throughput screening methodologies for biosensor engineering, optogenetics, genetic circuits, high throughput single-cell sequencing and gene expression analysis, neuroinflammation, neurodegeneration and in vivo applications. We anticipate that these new biosensors and modulators will be transformative tools for both neuroscience and immunology research and be a powerful therapeutic by enabling noninvasive imaging of inflammatory responses and selective, inflammation-dependent immunomodulation with cell type specificity that can be used in deep brain structures, across large areas, in behaving animals.