Available Technology
Low voltage nanowire-based electroporation of cells
Technology:
Delivering biomolecules to cells by electroporation using nanowire forests
Markets Addressed
Electroporation, a method of making cell membranes porous by subjecting the cells to a voltage, is widely used to deliver diverse biomolecules into cells. But the existing electroporation techniques demand very high operation voltage (>1000 V), which makes instrumentation complicated and expensive, and causes cell death due to arcing events. Also, current methods do not work well with small cells, such as neurons. This invention, which uses nanowire forests for electroporation, can solve many of these issues, and lead to a variety of potential implementations:
1. High-throughput delivery of biomolecules (e.g., DNA, RNA or protein) into all types of cells, large cells like E. coli and yeast cells, as well as small ones like stem cells and neurons.
2. Site-specific biomolecular delivery within networks or systems of interacting cells. By constructing individually addressable nanowire electrodes, different sites of interest in the same cell or different cells within a network can be located and excited to intake biomolecules.
3. Different molecules can be repetitively or sequentially delivered to the same cells at different time points. These cells can remain on the nanostructured electrode between electroporations and still preserve their viability.
Innovations and Advantages
Professor Hongkun Park’s lab has created a nanowire array structure for electroporation. This platform consists of vertically aligned silicon nanowire arrays coated with electrically conductive material such as Au, Pt, or Ag. Individually addressable nanowire electrodes are also available via semiconductor processing. Target cells are cultured a top the nanowire electrodes that partially penetrate into the cells. Electroporation is achieved by applying an electric potential across the nanowire electrode and a metal electrode, both of which are immersed in a bath solution containing the biomolecule of interest.
Due to the dramatically enhanced electric field strength between the tip of nanowire and metal electrode, only a few volts are needed to deliver biomolecules into target cells. The delivery efficiency can be optimized by finely tuning the nanowire density, aspect ratio and radius of curvature. The nanowire platform has been successfully demonstrated on both easy to transfect cells (HEK cells) and hard to transfect cells (neurons). When delivering a membrane-impermeable dye molecules into HEK293 cells, we have reached a greater than 97% efficiency and more than 85% cell viability. Comparable efficiency and viability have also been achieved on neurons. In contrast, the commercial electroporation systems typically report ~70-90% efficiency for easy to transfect cells, and ~20-50% for hard to transfect cells.
Additional Information
Intellectual Property Status: U.S. Provisional Application filed March 14, 2011
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Inventor(s):
Jorgolli, Marsela
Park, Hongkun
Robinson, Jacob
Shalek, Alexander K.
Sutton, Amy A.
Categories:
For further information, please contact:
Mick Sawka, Director of Business Development
(617) 496-3830
Reference Harvard Case #4095