Pharmacodynamic approach to study the gene transfer process employing non-viral vectors.
In the present work we set out to apply pharmacodynamic concepts derived from dose-response curves (Potency and Efficacy) to characterize the gene transfer efficiency of a vector:DNA complex. We employed two widely used vectors, the cationic lipid DOTAP (N,N, N-trimethyl 1-2-3-bis (1-oxo-9-octa-decenyl)oxy-(Z, Z)-1-propanaminium methyl sulfate) and the cationic polymer PEI (polyethylenimine, 800 kDa) to transfect several constructions of the green fluorescent protein cDNA. The analysis of dose-response curves indicated that in all cases the goodness-of-fit was > 0.99. Potency is a measure that provides information on gene activity per amount of DNA. Efficacy is a measure of maximum gene expression achievable using a specific vector:DNA complex, and depends on both the intrinsic efficacy of the gene (evaluated using different vectors to transfer the same gene construct) and on vector efficacy in DNA delivery (evaluated using a single vector to deliver different gene constructs). The results suggest that Potency and Efficacy are objective parameters for describing and comparing the goodness of vectors, as well as the intrinsic efficacy of a given gene construct. Furthermore, they are useful tools that may contribute to a better understanding of the mechanistic gene transfer process of each vector.
Aliño
S F
SF
Departamento de Farmacología, Facultad de Medicina y Odontología, Universitat de València, Valencia, Spain. alino@uv.es
Escrig
E
E
Revert
F
F
Guillem
V M
VM
Crespo
A
A
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004247:DNA / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005229:Fatty Acids, Monounsaturated / Q000008:administration & dosage; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D000644:Quaternary Ammonium Compounds / Q000008:administration & dosage; D014407:Tumor Cells, Cultured
D000900:Anti-Bacterial Agents; D005229:Fatty Acids, Monounsaturated; D005839:Gentamicins; D000644:Quaternary Ammonium Compounds; D004247:DNA; C010680:antibiotic G 418; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane