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

PFI:AIR - TT：细菌高通量遗传转化平台

• 批准号: 1640678
• 负责人: Cullen Buie
• 金额: $ 20万
• 依托单位: Massachusetts Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2018-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1640678&HistoricalAwards=false
• 关键词: PFI; AIR; TT; Platform; Throughput

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.

该 PFI：AIR 技术翻译项目专注于翻译下一代微流体遗传转化技术，以满足高通量生成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。 这种新颖的遗传转化系统非常重要，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。该遗传转化平台处理微生物的速度将比现有技术快近一百倍。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（使用脉冲电场进行细胞透化）是将 DNA 构建体传递到微生物中以用于基因工程和合成生物学应用的有效方法。标准电穿孔方案涉及平行板比色皿，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次实验（例如不同的 DNA 构建体、细胞类型、缓冲液成分），速度慢、劳动密集型且昂贵。该项目旨在开发一个原型高通量平台，用于使用微流体流通电穿孔进行细菌遗传转化。该原型设备将在 1) 生物工程化学品的大规模生产中运行，2) 以发现模式运行，以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制造的新型生物工程化学品的开发。参与该项目的人员，包括研究科学家、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、与导师互动以及成立公司将技术商业化，获得创新、创业和技术转化的经验。该项目利用麻省理工学院风险指导服务和麻省理工学院创新计划来指导这项技术转化工作中从研究发现到商业现实的商业化工作。

项目成果

Toward establishing model organisms for marine protists: Successful transfection protocols for Parabodo caudatus (Kinetoplastida: Excavata): Protist transfection

建立海洋原生生物模型生物：Parabodo caudatus（动质体：Excavata）的成功转染方案：原生生物转染

• DOI: 10.1111/1462-2920.13830
• 发表时间: 2017-09
• 期刊: Environmental Microbiology
• 影响因子: 5.1
• 作者: Gomaa, Fatma;Garcia, Paulo A.;Delaney, Jennifer;Girguis, Peter R.;Buie, Cullen R.;Edgcomb, Virginia P.
• 通讯作者: Edgcomb, Virginia P.