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

这个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，以满足高通量一代转基因微生物的需求。该系统将使代谢工程师能够更快地开发微生物，用于生产各种生物工程化学品和材料。这种新型的基因转化系统非常重要，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。这个基因转化平台处理微生物的速度将比目前的技术快近100倍。该项目解决了以下技术差距，因为它从研究发现转化为商业应用。电穿孔是一种利用脉冲电场进行细胞渗透的方法，是一种将DNA结构体传递到微生物体内的有效方法，可用于基因工程和合成生物学。标准的电穿孔方案包括平行板比色皿，将细胞和DNA样品暴露在均匀的电场中。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结构，细胞类型，缓冲液组成），这是缓慢的，劳动密集型的，昂贵的。本项目旨在开发一种原型高通量平台，用于利用微流体通过电穿孔进行细菌的遗传转化。该原型装置将用于1)大规模生产生物工程化学品，2)以发现模式确定基因工程的最佳转化方案。这种可扩展的技术将加速以可再生方式生产的新型生物工程化学品的开发。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，与导师互动以及建立公司以实现技术商业化，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，指导这项技术从研究发现到商业现实的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.