IDBR: Type A - A"Smart" Electroporation Device for Controlled Permeabilization and Molecular Delivery

IDBR：A 型 - 用于受控透化和分子传递的“智能”电穿孔装置

• 批准号: 1353918
• 负责人: David Shreiber
• 金额: $ 42.57万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1353918&HistoricalAwards=false
• 关键词: IDBR; Type; Smart; Electroporation; Device

This award is being made jointly by two Programs- (1) Instrument Development for Biological Research, in the Division of Biological Infrastructure (Biological Sciences Directorate), and (2) Nano-Biosensing, in the Division of Chemical, Bioengineering, Environmental and Transport Systems (Engineering Directorate). Non Technical Description:Electroporation is a method frequently used to deliver genetic material (DNA and RNA) into cells that have proven to be difficult to transfect, such as stem cells, which are important both as a research model to understand development and disease and as a cell source for regenerative medicine. Cells are exposed to a brief, high strength electric field, which causes the cell membrane to become permeable temporarily, allowing transport of the molecules into the cell before the membrane reseals. Finding the right field strength has been done exclusively by trial-and-error, and even after the process is "optimized" for a cell type, there is natural and significant variability among those cells, leading to cell death or lack of delivery. The proposed project addresses this gap in technology by developing a "smart" electroporation system that recognizes the state of permeability of each cell and dynamically controls the pulse to prevent over- exposure to high strength fields while still allowing molecular delivery. The end result will be a technology that is easy to use, reproducible, and robust; and of value to laboratories conducting basic research as well as those in the biotechnology sector. A team of four scientists will conduct this project along with graduate and undergraduate students. K-12 outreach activities are planned to encourage students to develop interests in STEM. Technical Description:The instrument to be developed will monitor changes in the electrical characteristics of a cell as it becomes permeabilized, and modulate the applied electric field to safely and efficiently deliver the molecular payload. The devices will be validated across a range of molecule types, including small organic compounds, small interfering RNA (siRNA), and plasmid DNA that range in size from 1kDa to 1000 KDa. The instrument?s performance will be benchmarked against current state-of-the-art commercial technology for delivery of the range of molecules into NIH 3T3 fibroblasts, which are a frequent model cell used in cell biology, and into human lymphoblastoid cells, which are difficult to transfect but extremely valuable as a platform to generate induced pluripotent stem cells. Dissemination of the research, and the "smart" electroporator, will be achieved by presentations at national conferences and meetings within the scientific disciplines of engineering and microfabrication, as well as at experimental biology meetings to reach the end-users. Collaborations and partnerships with entities at Rutgers, such as the NJ Stem Cell Training Course and The Rutgers University Cell & DNA Repository Infinite Biologics, will be utilized to solicit feedback from the biomedical research community in academic, government, and industry sectors and demonstrate the instrument's capabilities, and the Center for Innovative Ventures of Emerging Technology, and the Office of Technology and Commercialization, to commercialize the technology and prepare for its distribution.

该奖项由两个计划（1）生物学研究工具开发，生物基础设施（生物科学局）共同颁发，以及（2）纳米生物传感器在化学，生物工程，环境和运输系统（工程局）的纳米生物传感器。非技术描述：电穿孔是一种通常用于将遗传物质（DNA和RNA）传递到已被证明很难转染的细胞中的方法，例如干细胞，它们是了解发育和疾病的研究模型，并且是再生医学的细胞来源。细胞暴露于短暂的高强度电场，这会导致细胞膜暂时变得渗透，从而使分子在膜重新密封之前将分子传输到细胞中。找到合适的场强度是通过试用和错误进行的，即使在该过程“优化”细胞类型后，这些细胞之间也存在自然而显着的可变性，从而导致细胞死亡或缺乏递送。 拟议的项目通过开发一个“智能”电穿孔系统来解决这一差距，该系统识别每个细胞的渗透性状态并动态控制脉冲以防止过度暴露于高强度场，同时仍允许分子递送。最终结果将是一项易于使用，可重现和健壮的技术。以及对进行基础研究以及生物技术领域的实验室的价值。由四个科学家组成的团队将与研究生和本科生一起进行该项目。计划开展K-12外展活动，以鼓励学生发展STEM的兴趣。 技术描述：要开发的仪器将在透化时监视细胞的电气特性的变化，并调节施加的电场以安全有效地传递分子有效载荷。这些设备将在各种分子类型的范围内进行验证，包括小有机化合物，小型干扰RNA（siRNA）和质粒DNA的大小从1KDA到1000 kDa。该仪器的性能将针对当前最新的商业技术进行基准测试，以将分子范围传递到NIH 3T3成纤维细胞中，该仪表是一种经常用于细胞生物学的模型细胞，并将其用于人类淋巴细胞细胞，这些细胞很难转染，但它是产生诱导诱导的Pluripotent pluripotent pluroipotent的平台的极其有价值的。这项研究的传播以及“智能”电氧化器将通过在工程和微观加工科学学科以及实验生物学会议上的国家会议和会议上的演讲来实现，以达到最终用户。 Collaborations and partnerships with entities at Rutgers, such as the NJ Stem Cell Training Course and The Rutgers University Cell & DNA Repository Infinite Biologics, will be utilized to solicit feedback from the biomedical research community in academic, government, and industry sectors and demonstrate the instrument's capabilities, and the Center for Innovative Ventures of Emerging Technology, and the Office of Technology and Commercialization, to commercialize the technology and prepare for its 分配。