Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizer.
A plasmid expressing the beta-galactosidase enzyme was used to transfect Vero cells in order to evaluate the efficiency of a liposome-mediated transfection by circular and linear DNA. The results obtained showed a low rate of transfection by linear DNA:liposome complexes. To explore whether the structure of the complexes was interfering with the transfection, atomic force microscopy (AFM) was used. It has confirmed the difference between the linear and circular condensates: whereas the circular DNA:liposome complexes presented compact spherical or cylindrical structures of about 100-800 nm, the linear DNA showed pearl necklace-like structures, with pearls varying from 250 to 400 nm. On the basis of the theory proposed by Kuhn et al. (1999), low concentrations of cationic amphihile were used to neutralize or reverse the DNA charge in order to improve the transfection efficiency of the linear DNA. Using this method, we were able to obtain the expression of the transgene without an associated toxicity observed with the linear DNA liposome delivery.
von Groll
Andrea
A
Departamento de Patologia Clínica Veterinaria, Faculdade de Veterinaria, and Departamento de Física Teórica, Instituto de Física, Universidade Federal do Rio Grande do Sul, Campus do Vale, Porto Alegre, RS, Brazil.
Levin
Yan
Y
Barbosa
Marcia C
MC
Ravazzolo
Ana P
AP
D000818:Animals; D002412:Cations / Q000737:chemistry; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000379:methods; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity; D014162:Transfection / Q000379:methods*; D014709:Vero Cells; D001616:beta-Galactosidase / Q000737:chemistry / Q000378:metabolism
D002412:Cations; D008055:Lipids; D008081:Liposomes; D004247:DNA; D001616:beta-Galactosidase