Considerations for the Design of Improved Cationic Amphiphile-Based Transfection Reagents

Abstract

Cationic amphiphiles (cytofectins) are widely used for the transfection of cultured cells, and may become useful for the development of genetic medicines. Although fundamental research focused on clarification of physicochemical structure/biologic function correlations has been limited, general principles relating to optimization of cytofectin structure are beginning to emerge. We review these general conclusions and suggest considerations which may assist in the development of improved compounds. Data which relates to formulation is also discussed. The formulation studies address the tendency of high concentration cytofectin:polynucleotide complexes to precipitate. From these observations we postulate that a thermodynamically stable product can be formed by sonication with heating of cytofectin:polynucleotide complexes, and that this process reduces the kinetically driven aggregation and precipitation which currently complicates many in vivo studies.