I-Corps: Kytopen: Expanding the language of biology with pulsed electric fields

I-Corps：Kytopen：用脉冲电场扩展生物学语言

• 批准号: 1562925
• 负责人: Cullen Buie
• 金额: $ 5万
• 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: 2015
• 资助国家: 美国
• 起止时间: 2015-10-15 至 2017-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1562925&HistoricalAwards=false
• 关键词: Corps; Kytopen; Expanding; language; biology

The potential to solve many of mankind's most pressing challenges including the needs for alternative fuels, enhancing oil recovery, and even treating cancer could involve engineering the numerous bacteria currently beyond one's reach. There are many types of bacteria that can be grown in the lab but their potential impact on people's lives has not been realized. New tools are needed to unlock bacteria?s true potential to solve many challenges of interest to mankind. Creating tools to accelerate discovery of new cancer treatments, alternative fuels, or low cost biomaterials will significantly impact human life. Population growth and increasing lifespans are putting ever increasing demands on the environment, driving need for alternative sources of fuels, food, and medicines. Bacteria have been a bit neglected as sources for innovation; however, they already have the internal machinery to help do industrial processes in a fast and efficient manner.Pulsed electric fields (i.e. electroporation) can be used to deliver genetic material into many types of cells including bacteria. The goal of this proposal is to substantially expand the successful application of pulsed electric fields to bacteria and enable discovery of new applications in genetic engineering and synthetic biology. A major limitation of synthetic biology is the inability to incorporate genetic material into many bacteria due to the challenge of permeating the cell envelope while maintaining high cell viability. This team's goal is to reduce the time required to develop tools to genetically manipulate bacteria from months or even years to days. This I-Corps team has developed a proof-of-concept microfluidic device to enable the characterization of the critical electric field for bacterial electroporation under specific experimental conditions in a single experiment. The team is devising a technique to test several thousand unique conditions within a single day, spanning the entire parametric space. This will be done using a microfluidic platform, or several in parallel, to test small aliquots of cells at varying experimental conditions. Therefore, the proposed microfluidic system enables quantification of the critical electroporation parameters in a single experiment, which would otherwise require months of experimentation. The proposed innovation will allow two significant advancements: 1) the ability to quickly determine high yield electroporation conditions and 2) the ability to utilize many of the more than 10,000 bacterial strains that have been heretofore intractable or difficult to genetically manipulate.

解决人类许多最紧迫的挑战的潜力，包括对替代燃料的需求，提高石油采收率，甚至治疗癌症，可能需要对目前人们无法企及的大量细菌进行改造。有许多类型的细菌可以在实验室中培养，但它们对人们生活的潜在影响还没有意识到。需要新的工具来解锁细菌吗？S真正有潜力解决许多人类感兴趣的挑战。创造工具来加速发现新的癌症治疗方法、替代燃料或低成本生物材料将对人类生活产生重大影响。人口的增长和寿命的延长对环境提出了越来越高的要求，推动了对替代燃料、食品和药品的需求。细菌作为创新的来源一直被忽视；然而，它们已经拥有帮助快速高效地进行工业过程的内部机械。脉冲电场(即电穿孔)可以被用来将遗传物质输送到包括细菌在内的许多类型的细胞中。这项提议的目标是大幅扩大脉冲电场对细菌的成功应用，并使其能够在遗传工程和合成生物学中发现新的应用。合成生物学的一个主要限制是无法将遗传物质结合到许多细菌中，这是因为在保持高细胞活力的同时渗透到细胞膜的挑战。该团队的目标是将开发基因操作细菌工具所需的时间从几个月甚至几年减少到几天。这个i-Corps团队开发了一种概念验证微流控设备，能够在单个实验中表征特定实验条件下细菌电穿孔的临界电场。该团队正在设计一种技术，在一天内测试数千种独特的条件，跨越整个参数空间。这将使用一个微流控平台，或几个并行的平台，在不同的实验条件下测试小等份的细胞。因此，拟议的微流控系统能够在一次实验中量化关键的电穿孔参数，否则这将需要几个月的实验。这项拟议的创新将带来两个重大进步：1)快速确定高产电穿孔条件的能力；2)利用迄今为止难以解决或难以进行基因操作的1万多个细菌菌株中的许多菌株的能力。