STTR Phase I: Polymer electrosonic actuation microarray: Low-cost tool for transfection of biological cells

This Small Business Technology Transfer (STTR) Phase I project will prove the technical and commercial feasibility of a single-use polymer microarray for electromechanical transfection of biological cells. Alternative gene transfer solutions are needed to access the potential held in biopharmaceuticals, gene therapy, and stem cell research. Although physical methods for gene transfer (e.g., electroporation) have demonstrated improved (over conventional chemical-mediated techniques) treatment outcomes in difficult-to-transfect primary and stem cells, control and uniformity of treatment remain inadequate. Further, incremental improvements in performance of cuvette-based electroporation systems have only been realized through corresponding increases in system complexity and cost. One-by-one ejection of cells through cell-sized orifices has been found to promote cell membrane poration and DNA delivery into cells by imposing an identical and carefully controlled electromechanical environment on each individual cell. Unfortunately, a relatively high material cost and limitations on achievable treatment conditions (due to constraints on the geometry of microarrays manufactured using standard silicon micromachining techniques), inhibit practical implementation of existing silicon-based microarrays. The innovative design and optimization of the polymer microarray under this STTR Phase I project will yield a low-cost system capable of generating the mechanical stress field needed to achieve improved treatment outcomes in difficult-to-transfect cells. The broader impact/commercial potential of this project is to enable effective and economical transfection of difficult-to-transfect primary and stem cells used for a variety of research and therapeutic applications in the Life Sciences. Effective delivery of genes, drug molecules, imaging agents, peptides, antibodies, and enzymes into living cells is critical to applications ranging from the treatment of human disease through introduction of DNA to the investigation of basic cellular function through single molecule imaging; yet, intracellular delivery and transfection remain difficult tasks. While efficiencies of greater than 90% are common in basic research applications that use chemical or physical methods to transfect laboratory established and maintained ("easy") cell lines, efficiency can drop to 10% or lower for "difficult" cells. Refinements of physical methods (e.g., electroporation) have achieved incremental performance improvements; however, no system currently on the market meets all end-user requirements for efficiency, viability, functionality, and cost. The novel approach to transfection, which is the subject of this STTR Phase I project, promises to improve treatment efficacy through innovative use of multiple gene transfer techniques simultaneously, while better addressing end user needs by providing a cost-effective transfection solution for difficult cells.