Collaborative Proposal: Engineering Bacterial Outer Membrane Vesicles for New Biotechnology Applications

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CBET 1264701/1264719DeLisa/ChenOuter membrane vesicles (OMVs) are nanoscale proteoliposomes derived from Gram-negative bacteria such as Escherichia coli. It is now firmly established that bacteria can be engineered to produce recombinant proteins in the different micro-compartments (i.e., lumen, membrane, outer surface) of an OMV. This capability has been widely exploited for vaccine development; however, very little else has been done to harness the full potential of OMVs. This project seeks to extend OMVs for other untapped applications typically performed by synthetic polymer-based vesicles. This will be achieved by expanding the range and complexity of biomolecular structures that can be functionally produced in the lumens, in the membranes, and on the surfaces of OMVs. The intellectual merit of the proposed research is the development of a versatile array of tools to independently or simultaneously embed functions into the different compartments of OMVs using standard molecular biology techniques rather than the tedious and multiple-step syntheses and conjugations associated with polymer-based vesicle systems.The PIs will leverage their combined expertise to create synthetic OMVs with the potential to impact many scientific areas, ranging from cell-specific targeting, gene/protein delivery, vaccine development, cascading enzyme reactions, biosensing and bioremediation. The proposed research is also highly interdisciplinary in nature. Graduate students participating in this project will gain an integrated perspective of the important interfaces and synergies connecting biochemistry, microbiology, modern genetics, synthetic biology and bioengineering. These students will also gain an appreciation for translating fundamental discoveries into practical technologies. The broader impacts of the project include an extended outreach program for disadvantaged students from New York City who face significant economic disadvantages and a Homeschoolers Day Program that seeks to work with local homeschool families.Due to the interdisciplinary nature of the project, this award by the Biotechnology, Biochemical, and Biomass Engineering Program of the CBET Division is co-funded by the Biomaterials Program of the Division of Materials Research.