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

这项小企业创新研究（SBIR）第一阶段项目的广泛影响是开发一种新的系统，以提高转基因生物的安全性和生产力。转基因生物使新的活疗法、农产品和生物基消费品成为可能。然而，一个关键的挑战是缺乏可用的工具来轻松控制转基因生物的行为。该提案将开发一种技术，通过将转基因生物的生存与特定控制化学品的存在联系起来，来确保遏制转基因生物。核心技术是基于经过改造的细胞，这些细胞可以根据特定的化学信号改变它们的行为，而这个项目的目标是扩大可以由化学输入控制的蛋白质组合（以及行为）。这解决了人们对转基因生物的使用和可能产生的未知后果的担忧。拟议的项目将建立一个化学控制的基本基因组合。这将通过蛋白质工程来实现，同时构建12,000个蛋白质文库，涵盖200个不同的必需基因。利用靶向基因组整合，该文库将被整合到大肠杆菌的基因组中。使用阳性和阴性（化学）选择对文库进行表型表征，然后进行下一代DNA测序，以揭示单个蛋白质设计的功能（致死、活或有条件）。该数据集将阐明结构、序列和功能之间的关系，用于化学控制蛋白质的设计。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.