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.
|Effective start/end date||8/1/14 → 7/31/17|
- National Science Foundation (National Science Foundation (NSF))