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

CBET 1264701/1264719 DeLisa/ChenOuter membrane vesicles（OMV）是来自革兰氏阴性细菌如大肠杆菌的纳米级蛋白脂质体。现在已经确定，可以对细菌进行工程改造，以在不同的微区室中产生重组蛋白（即，腔、膜、外表面）。这种能力已被广泛用于疫苗开发;然而，在利用OMV的全部潜力方面，几乎没有做过其他工作。该项目旨在将OMV扩展到通常由合成聚合物囊泡执行的其他未开发应用。这将通过扩大生物分子结构的范围和复杂性来实现，所述生物分子结构可以在管腔中、膜中和OMV表面上功能性地产生。这项研究的智力价值在于开发一系列多功能工具，使用标准分子生物学技术独立或同时将功能嵌入到OMV的不同隔室中，而不是与基于聚合物的囊泡系统相关的繁琐和多步合成和缀合。PI将利用他们的综合专业知识来创造具有影响许多科学领域潜力的合成OMV，范围从细胞特异性靶向、基因/蛋白质递送、疫苗开发、级联酶反应、生物传感和生物修复。拟议的研究也是高度跨学科的性质。参与该项目的研究生将获得连接生物化学，微生物学，现代遗传学，合成生物学和生物工程的重要接口和协同作用的综合视角。这些学生还将获得将基本发现转化为实用技术的赞赏。该项目更广泛的影响包括为纽约市面临重大经济困难的弱势学生提供的扩展外展计划，以及寻求与当地家庭学校家庭合作的家庭学校日计划。由于该项目的跨学科性质，生物技术，生物化学，CBET部门的生物质工程项目由材料研究部门的生物材料项目共同资助。

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.