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

该奖项由两个项目联合颁发-（1）生物基础设施司（生物科学理事会）的生物研究仪器开发，以及（2）化学，生物工程，环境和运输系统司（工程理事会）的纳米生物传感。电穿孔是一种经常用于将遗传物质（DNA和RNA）递送到已被证明难以转染的细胞（如干细胞）中的方法，干细胞作为了解发育和疾病的研究模型以及再生医学的细胞来源都很重要。细胞暴露于短暂的高强度电场，这导致细胞膜暂时变得可渗透，允许在膜重新密封之前将分子运输到细胞中。找到正确的场强完全是通过试错法完成的，即使在该过程针对细胞类型进行了“优化”之后，这些细胞之间也存在自然和显著的变异性，导致细胞死亡或缺乏递送。 拟议的项目通过开发一种“智能”电穿孔系统来解决这一技术差距，该系统可以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，同时仍然允许分子输送。最终的结果将是一种易于使用、可重复和强大的技术;对进行基础研究的实验室以及生物技术部门的实验室都有价值。一个由四名科学家组成的团队将与研究生和本科生沿着进行这个项目。计划开展K-12外展活动，鼓励学生培养STEM兴趣。 技术说明：待开发的仪器将监测细胞透化时电特性的变化，并调节所施加的电场，以安全有效地递送分子有效载荷。这些设备将在一系列分子类型中进行验证，包括小分子有机化合物、小干扰RNA（siRNA）和质粒DNA，其大小范围为1 kDa至1000 KDa。乐器？的性能将以目前最先进的商业技术为基准，用于将一系列分子递送到NIH 3 T3成纤维细胞和人类淋巴母细胞中，NIH 3 T3成纤维细胞是细胞生物学中常用的模型细胞，人类淋巴母细胞难以分离，但作为产生诱导多能干细胞的平台极具价值。将通过在工程和微加工科学学科的全国性会议和会议上以及在实验生物学会议上向最终用户介绍研究成果和“智能”电穿孔仪。与罗格斯大学实体的合作和伙伴关系，如新泽西州干细胞培训课程和罗格斯大学细胞DNA库无限生物制品，将用于征求学术界，政府和工业部门的生物医学研究界的反馈，并展示仪器的能力，以及新兴技术创新风险投资中心，以及技术和商业化办公室，将技术商业化并为其分配做准备。

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.