Construction of Cationic Virosome Derived from Vesicular Stomatitis Virus as a Promising Candidate for Efficient Gene Delivery to the Central Nervous System

Introduction: Nowadays, one of the barriers of gene therapy in the treatment of the CNS diseases is the lack of proper and safe carrier sys tems to cross the blood brain barrier (BBB).Virosomes are virus like particles which can be used in brain if made from neurotropic viruses.The aim of the s tudy was to cons truct cationic virosomes derived from vesicular s tomatitis virus using dialyzable short chain phospholipid (DCPC) and cationic lipid (DOTAP) invitro. Materials and Methods:The vesicular s tomatitis virus was propagated in Vero cell line.Subsequently, the harves ted virus was concentrated and purified using ultrafiltration and ultracentrifugation and finally, the virosome was synthesized by DCPC detergent and the addition of cationic lipid.Particle size dis tribution of virosome nanoparticles, cellular cytotoxicity and glycoprotein of vesicular s tomatitis virus (VSV-G) were determined by measuring dynamic light scattering using zetasizer, MTT assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively.Results: The harves ted viruses were concentrated and purified by ultrafiltration and ultracentrifugation and the final concentration was 0.8 mg/ml.The cationic virosome mean size was 186.6 nm and, the cell viability was significantly decreased after 48 hours of treatment with different concentrations of virosome compared to the control group.The VSV-G protein with molecular weight of 63 kDa was approved by SDS-PAGE.Conclusion: The use of DCPC is an efficient method for solubilization and recons truction of vesicular s tomatitis virus envelope and does not alter the surface VSV-G.Due to the VSV-G protein and its wide range cell tropism, this cationic virosome can also be a promising candidate for crossing the BBB in order to efficient gene delivery and therapy of CNS diseases.