Non-covalent association of folate to lipoplexes: a promising strategy to improve gene delivery in the presence of serum.
The success of gene therapy depends on the efficient delivery of therapeutic genes into target cells in vitro and in vivo. Non-viral vectors, such as cationic liposome-DNA complexes (lipoplexes), have been used for numerous gene delivery applications, although their efficacy is still limited, particularly when compared to that of viral vectors. In this work, we assessed the efficacy of a new gene delivery system generated by non-covalent association of folate to lipoplexes (FA-associated lipoplexes) in two different cancer cell lines (SCC-VII and TSA cells). Association of FA with liposomes composed of DOTAP and cholesterol, and subsequent complexation with DNA greatly increased transfection efficiency above that obtained with plain lipoplexes in both cell lines. The addition of 40μg of FA to lipoplexes was optimal for transfection and allowed to overcome the inhibitory effect induced by the presence of serum. Notably, the biological activity of the FA-associated complexes was even significantly improved under these conditions. Transfection activity mediated by FA-associated lipoplexes was compared with that by FA-conjugated lipoplexes, and the results showed that electrostatic association of FA to the lipoplexes led to considerably higher levels of biological activity than that involving covalent coupling of FA. Moreover, FA-associated lipoplexes confer greater DNA protection than FA-conjugated lipoplexes. To our knowledge, this is the first study reporting the characterization and application of FA-associated lipoplexes in gene delivery and showing their greater efficacy than that of FA-conjugated lipoplexes. Overall, the results obtained in the present work constitute a strong indication that the developed FA-associated lipoplexes are promising candidates for in vivo gene delivery.
Center for Neuroscience and Cell Biology of Coimbra, University of Coimbra, Coimbra, Portugal.
Sónia
S
Duarte
Henrique
H
Faneca
Maria C Pedroso
MC
de Lima
D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D005492:Folic Acid / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D009369:Neoplasms / Q000628:therapy; D044967:Serum / Q000378:metabolism*; D055672:Static Electricity; D014162:Transfection / Q000379:methods*
D008081:Liposomes; D049452:Green Fluorescent Proteins; D004247:DNA; D005492:Folic Acid