(23b) Optimization of Nanoparticle Delivery Platform for mRNA

The delivery of functional gene into cells to replace mutated genes holds great potential for treatment of many type diseases. In this line of investigation, mRNA therapy has gathered great interest because of its potential role for the transient modification of immune cells for treatment of an array of diseases. However, the main obstacle preventing routine mRNA immune cell transfection is the lack of a high viability, high efficiency, high throughput delivery system. In this study, we propose the use of a physical mixture of sterilized hollow gold nanoshells (HGN), that can be irradiated with picosecond pulsed near infra red light to induce vapor nanobubbles around the HGN which rupture the endosomes, thereby releasing the mRNA to the cell cytoplasm, where the mRNA can produce the proteins of interest. In Summary, we present extensive analysis into HGN sterilization techniques to minimize degradation of mRNA thus allowing the prolonged availability of mRNA for efficient translation as well as increased transfection efficiency in Jurkat and HEK-273 cells. The HGN/mRNA mixture had successful cell transfection with no negative effects on cell viability post transfection. Experiments are underway to determine the stability of genetic material in biological environment and cell extracts using Fluorescence Correlation Spectroscopy. Furthermore, we will investigate the use of Pluoronics to increase structural stability and resealing of cell membrane after irradiation with infrared light.

This Study is funded by the Heltzer Chair at U of M and Regenerative Medicine Minnesota.