The investigators propose a novel method to pharmaceutically engineer solid nanoparticles in the size range of 5 to 50 nm containing plasmid DNA fornon-viral gene therapy applications. The method potentially overcomeslimitations of current non-viral gene delivery technology which uses positively-charged lipids or polymers to form larger, unstable complexeswith negatively-charged plasmid DNA in the size range of 100 to 1000 nm. Further, using the microemulsion precursor method to engineer solid nanoparticles, well-defined and uniform solid nanoparticles (5 to 50 nm)may be spontaneously and reproducibly made without the use of expensive or potentially damaging techniques involving high-torque mechanical mixing, microfluidization, homogenization, or milling. The proposed method involves the spontaneous formation of novel microemulsions wherein the dispersed droplets, which are typically 5 to 50 nm in diameter, are precursors for the formation of solid and stable nanoparticles. In addition to plasmid DNA, engineered nanoparticulate systems may contain many different materials for various medical and engineering applications such as nanomagnets and nanosensors.
The specific goal of the project is to demonstrate that targeted solid nanoparticles containing plasmid DNA can be engineered from novel microemulsion precursors. The project has three specific aims, 1) demonstrate that plasmid DNA can be incorporated into a stable microemulsion and that stable solid nanoparticles (5-50 nm) containing plasmid DNA can be made from the microemulsion precursor, 2) characterize the solid nanoparticles (i.e., size, surface charge and porosity, DNA release and stability) and demonstrate that the solid nanoparticles are stable in biological fluids and can express a transgene in-vitro, and 3) incorporate a cell-specific ligand onto the surface of the solid nanoparticles and demonstrate surface recognition of the targeted nanoparticles.
