There is a need to develop affordable and specific biosensors to defend against current and future biological threats. Biological methods of detection have been evolved by nature to detect molecules at the micro-scale with flexible specificity and with downstream effectors. Advances in cell-free technology allow for deployment of biosensors on pH-strip-type paper, an affordable, robust, disposable, and highly portable format for Field use. Synvitrobio and UW Madison are developing BioSENSE, a product built off of cell-free expression to produce allosteric transcription factor (aTF) biosensors to DoD-relevant molecules with turnaround of 2-months or less. BioSENSE combines computational aTF design with high-throughput cell-free expression to rapidly test design iterations for activity. In Phase I, we demonstrated the development and use of BioSENSE to sense model molecules using model and engineered aTF biosensors. In Phase II, we propose scaling BioSENSE to develop biosensors towards 10 DoD-defined molecules of interest across 5 classes (pathogenic markers, environmental pollutants, explosives, human performance markers, and toxins). The identified biosensors can be implemented on paper for deployable detection, or can be used within engineered cells.
