PFI:AIR - TT: A Platform for High Throughput Genetic Transformation of Bacteria

该PFI：AIR技术翻译项目的重点是翻译下一代微流体遗传转化技术，以满足高通量生产转基因微生物的需求。该系统将允许代谢工程师更快地开发用于生产各种生物工程化学品和材料的微生物。 这种新的遗传转化系统很重要，因为合成生物学和基因工程领域目前受到用外源DNA重新编程微生物的能力的限制。该项目将产生一个原型高通量遗传转化平台，以展示该系统的实用性。该基因转化平台将能够以比当前技术水平快近一百倍的速度处理微生物。该项目在从研究发现转向商业应用的过程中解决了以下技术差距。电穿孔，使用脉冲电场的细胞透化，是将DNA构建体递送到微生物中用于基因工程和合成生物学应用的有效方式。标准电穿孔方案涉及平行板比色皿以将细胞和DNA样品暴露于均匀电场。然而，测试不同的电穿孔条件涉及数百个实验（例如，不同的DNA构建体、细胞类型、缓冲液组成），这是缓慢的、劳动密集型的并且昂贵的。本项目旨在开发一个原型的高通量平台，用于使用微流体流通电穿孔进行细菌的遗传转化。该原型装置将用于1）生物工程化学品的大规模生产和2）以发现模式运行，以确定遗传工程的最佳转化方案。这种可扩展的技术将加速新的生物工程化学品的开发，这些化学品可以以可再生的方式生产。参与该项目的人员，研究科学家，博士后和本科生，将通过参与原型设计，客户访谈，商业计划竞赛，与导师合作以及建立公司将技术商业化来获得创新，创业和技术翻译经验。该项目与麻省理工学院创业指导服务和麻省理工学院创新计划合作，以指导从研究发现到商业现实的技术转化工作的商业化方面。

This PFI: AIR Technology Translation project focuses on translating next generation microfluidic genetic transformation technologies to fill the need for high throughput generation of genetically modified microorganisms. This system will allow metabolic engineers to more rapidly develop microorganisms for the production of various bioengineered chemicals and materials. This novel genetic transformation system is important because the fields of synthetic biology and genetic engineering are currently limited by the ability to re-program microorganisms with foreign DNA. This project will result in a prototype high throughput genetic transformation platform to demonstrate the utility of the system. This genetic transformation platform will be able to process microorganisms nearly one hundred times faster than the current state of the art. This project addresses the following technology gap(s) as it translates from research discovery toward commercial application. Electroporation, cell permeabilization using pulsed electric fields, is an efficient way to deliver DNA constructs into microorganisms for genetic engineering and synthetic biology applications. Standard electroporation protocols involve parallel plate cuvettes to expose cell and DNA samples to uniform electric fields. However, testing different electroporation conditions involves hundreds of experiments (e.g. varying DNA construct, cell type, buffer composition), which is slow, labor-intensive, and expensive. This project aims to develop a prototype high throughput platform for genetic transformation of bacteria using microfluidic flow-through electroporation. The prototype device will be operated in 1) large scale production of bioengineered chemicals and 2) in a discovery mode to identify optimal transformation protocols for genetic engineering. This scalable technology will accelerate the development of new bioengineered chemicals that can be manufactured in a renewable manner. Personnel involved in this project, research scientists, postdocs, and undergraduate students, will receive innovation, entrepreneurship, and technology translation experiences through participation in prototype design, customer interviews, business plan competitions, engaging with mentors, and establishing a company to commercialize the technology. The project engages the MIT Venture Mentoring Services and the MIT Innovation Initiative to guide commercialization aspects in this technology translation effort from research discovery toward commercial reality.