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

外膜囊泡（OMVs）是来源于革兰氏阴性菌（如大肠杆菌）的纳米级蛋白脂质体。现在已经确定，细菌可以在OMV的不同微室（即管腔、膜、外表面）中产生重组蛋白。这种能力已被广泛用于疫苗开发；然而，为了充分利用omv的潜力，几乎没有做过其他工作。该项目旨在将omv扩展到其他未开发的应用中，这些应用通常是由合成聚合物基囊泡完成的。这将通过扩大生物分子结构的范围和复杂性来实现，这些结构可以在管腔、膜和omv表面上功能性地产生。所提出的研究的智力价值在于开发了一系列通用的工具，可以使用标准的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且多步骤的合成和偶联。pi将利用他们的综合专业知识来创造合成的omv，这些omv有可能影响许多科学领域，包括细胞特异性靶向、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。拟议的研究在本质上也是高度跨学科的。参与该项目的研究生将获得一个综合的视角，了解生物化学、微生物学、现代遗传学、合成生物学和生物工程之间的重要接口和协同作用。这些学生还将学会如何将基础发现转化为实用技术。该项目更广泛的影响包括为纽约市面临严重经济劣势的弱势学生提供扩展外展计划，以及寻求与当地家庭合作的家庭学校日计划。由于该项目的跨学科性质，该奖项由CBET部门的生物技术，生化和生物质工程项目与材料研究部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.