SBIR Phase I: Development of a Novel Biocontainment/Biosafety Platform Using Synthetic Auxotrophs

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The broader impact of this Small Business Innovation Research (SBIR) Phase I project is the development of a novel system that will increase the safety and productivity of genetically modified organisms (GMOs). GMOs enablie novel live therapeutics, agricultural products, and bio-based consumer products. However, a key challenge is the lack of available tools to easily control the behavior of GMOs. This proposal will develop a technology that can be used to ensure containment of GMOs by tying their survival to the presence of specific control chemicals. The core technology is based on cells that have been engineered to alter their behavior in response to specific chemical signals, and the goal of this project is to expand the portfolio of proteins (and thus behaviors) that can be controlled by chemical inputs. This addresses concern surrounding the use and possible escape of GMOs that may have unknown consequences. The proposed project will establish a portfolio of chemically-controlled essential genes. This will be accomplished by protein engineering and simultaneously constructing 12,000 protein libraries spanning 200 different essential genes. Using targeted genome integration, this library of libraries will be integrated into the genome of Escherichia coli. Phenotypic characterization of the library using positive and negative (chemical) selections followed by next-generation DNA sequencing will be used to uncover the function (lethal, viable, or conditional) of individual protein designs. This dataset will illuminate the relationship between structure, sequence, and function for design of chemically-controlled proteins.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.