SBIR-STTR Award

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is the development a scalable, automated, genetic transformation platform that is 10,000X faster than the current state-of-the-art. The fields of synthetic biology and genetic engineering are currently limited by the ability to re-program microorganisms with foreign DNA. There have been significant advances in the synthesis of DNA, screening of genetically engineered microorganisms, and bioinformatics. However, the technology used to deliver DNA and perform genetic transformation has not advanced in a similar way. Phase I of this SBIR will result in a prototype high-throughput genetic transformation platform to demonstrate the utility of the system. This system will allow genetic engineers to more rapidly develop microorganisms for the production of bioengineered chemicals and materials. This SBIR Phase I project proposes to develop a high-throughput, automated platform for genetic transformation of bacteria using a proprietary flow-through electroporation technology that is fast, reliable, and scalable. A key step in genetic engineering of cells is to introduce the foreign DNA that re-programs the cell. Electroporation, cell permeabilization using pulsed electric fields, is the most efficient and widespread method to deliver DNA into microorganisms for this application. State-of-the-art electroporation involves cuvettes that expose the cells and DNA to uniform electric fields. However, this process is currently slow, labor-intensive, and expensive. The proposed technology can be automated by augmenting existing liquid handling robots, and, when operated in parallel, may improve the genetic transformation rate by up to 10,000X compared to current methods. This will represent a paradigm shift in areas dependent upon genetic transformation where DNA delivery using electroporation is currently a major bottleneck. Ultimately, the goal is to address the need for a high-throughput genetic transformation platform to accelerate innovation in synthetic biology.

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop a fast, efficient, and scalable cell engineering technology that is easily automated through integration with liquid handling robots. Currently, there is a bottleneck in the process of cell engineering, especially in the engineering of cells for discovery of new therapeutics. The field of delivery of genetic or other material to cells has not kept pace with advancements in genetic modification and high-throughput screening technologies. The proposed platform will offer an alternative to the time-consuming and labor-intensive methods of transfection including lentiviral transduction and cuvette-based electroporation, which are difficult to automate. Applications of cell engineering technology range from fundamental research in cell physiology to the discovery of new targets for cellular therapies. The platform will allow scientists and clinicians to more rapidly and reliably engineer immune and other cells for discovery of new therapeutic targets and therapeutics.The intellectual merit of this SBIR Phase II project will be to develop a scalable, automated, non-viral cell engineering platform with the potential to operate up to 10,000 times faster than conventional electroporation using high-throughput liquid handling. Using the core cell engineering technology developed in Phase I, the goal is to develop an automated protocol for gene transfection on a liquid handling robot compatible with 96 or 384 well plate technology. The first objective is to demonstrate the manufacturability of cell engineering devices for high-throughput cell engineering. Preliminary work in this area has shown that these devices can be injection molded, thus reducing cost while increasing the potential for production at scale. In the Phase II project, injection molded prototypes of the cell engineering devices will be developed to prove manufacturability and determine the cost to manufacture at scale (millions of parts per year). Second, there are several supplemental systems that must be integrated with a liquid handling apparatus to enable the proposed high-throughput cell engineering. Supplemental systems include a power source and power distribution manifold that interacts with each sample of the 96 or 384 well array. In this project, these systems will be integrated with the cell engineering devices and automated liquid handling robot. Third, the integrated system will be used to generate a large library of primary human T cell variants as proof-of-concept to demonstrate the potential for high-throughput cell engineering for therapeutic target discovery.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.