The research provided focuses on the development and application of cationic liposomes formed from mixtures of lecithin and dihexadecyldimethylammonium bromide (DDAB) for the purpose of efficient gene delivery. This study underscores the effectiveness of spontaneously formed cationic vesicles in transfecting plasmid DNA (pDNA) and siRNA into cells, showcasing a novel method for gene therapy. The experimental results reveal that by adjusting the DDAB to phospholipid ratio within these vesicles, their average size can be controlled within the range of 150-300 nm, with a ζ potential of +40 mV, indicative of their cationic nature. The ability of these vesicles to bind and form complexes with pDNA, referred to as lipoplexes, is thoroughly demonstrated through a variety of assays, including ζ potential measurement, isothermal titration calorimetry, and DNase I digestion assay, among others. A particularly significant finding is the binding enthalpy between pDNA and cationic liposome, calculated as -5.7 (±0.8) kJ/mol, emphasizing the favorable energetics of this interaction. Cellular uptake studies further confirm the vesicles' capability to efficiently deliver genetic material into MCF-7 and HeLa cells, with fluorescence microscopy showcasing successful transfection. Additionally, the study elaborates on the potential of these cationic vesicles in delivering siRNA for gene silencing, marking a significant step forward in the utilization of DDAB liposomes for gene delivery applications. This research holds profound implications for the field of gene therapy, presenting a scalable and effective method for the delivery of therapeutic genes and RNA into cells.
Shelar, S. B., Dey, A., et al. Spontaneous Formation of cationic vesicles in aqueous DDAB-Lecithin mixtures for efficient plasmid DNA complexation and gene transfection. ACS Applied Bio Materials. 2021, 4(8): 6005-6015.
For Research Use Only. Not For Clinical Use