A cationic lipid emulsion/DNA complex as a physically stable and serum-resistant gene delivery system.

Abstract

To develop a non-viral gene delivery system in the form of an oil-in-water (o/w) lipid emulsion. Cationic lipid emulsions were formulated with soybean oil, 1,2-dioleoyl-sn-glycero-3-trimethylammonium-propane (DOTAP) as a cationic emulsifier and other co-emulsifiers. The physical characteristics of the lipid emulsion and the emulsion/DNA complex were determined. The in vitro transfection efficiency of the emulsion/DNA complex was determined in the presence of up to 90% serum. The average droplet size and zeta potential of emulsions were ca. 180 nm and ca. +50 mV, respectively. Among the emulsions, a stable formulation was selected to form a complex with a plasmid DNA encoding chloramphenicol acetyltransferase. By increasing the ratio of emulsion to DNA. zeta-potential of the emulsion/DNA complex increased monotonously from negative to positive without any changes in the complex size. The complex was stable against DNase I digestion and an anionic poly-L-aspartic acid (PLAA). The complex delivered DNA into the cells successfully, and the transfection efficiency was not affected by complex formation time from 20 min to 2 h. More importantly, the cationic lipid emulsion facilitated the transfer of DNA in the presence of up to 90% serum. The cationic lipid emulsion/DNA complex has physical stability and serum resistant properties for gene transfer.