Cationic liposomes for gene transfection.
Cationic liposomes spontaneously interact with the negatively charged DNA to form a stable complex that promotes the gene transfer to cells. The mode of formation and the size of cationic liposomes/DNA complexes were investigated using the atomic force microscopy (AFM). Also the most important physical-chemical factors involved in cationic liposome-mediated gene transfection, e.g. size and lipidic composition, were evaluated through the transfection of complexes with different liposomes/DNA molar ratio into three types of cultured cells. Cationic liposomes, composed of a neutral lipid (phosphatidilcoline), a cationic lipid dimethyldioctadecylammonium bromide (DDAB), a co-lipid 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) and a phospholipid derivative of polyethylene glycol (DSPE-mPEG) at different molar ratio, were mixed with a plasmid pCMVbeta to form liposomes/DNA complexes. We have demonstrated that the complexes were made by complicated structures in which the liposomes tend to aggregate and the DNA is surrounded by lipidic material. In vitro transfection efficiency by liposomes/plasmid pCMVbeta complexes was found to depend on the kind of lipid associated in the liposomes and the liposomes/DNA mixing ratio. The importance of associating DOPE in cationic liposomes was confirmed; this co-lipid is able to improve the ability of cationic liposomes to transfect cells but in addition, the AFM images and the EtBr fluorescence experiments have suggested that this lipid can also play an important role to facilitate the formation of stable liposomes, which efficaciously protect the DNA by nuclease digestion.
Ruozi
Barbara
B
Department of Pharmaceutical Sciences, University of Modena and Reggio Emilia, Via Campi 183, 41100 Modena, Italy.
Forni
Flavio
F
Battini
Renata
R
Vandelli
Maria Angela
MA
D000818:Animals; D002412:Cations; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000008:administration & dosage; D004337:Drug Carriers; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007202:Indicators and Reagents; D008081:Liposomes / Q000138:chemical synthesis / Q000737:chemistry*; D051379:Mice; D018625:Microscopy, Atomic Force; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*
D002412:Cations; D004337:Drug Carriers; D007202:Indicators and Reagents; D008081:Liposomes; D004247:DNA