Biopolymers and cell. Volume 20. № 1-2. 25 - 32.
O. K. Toporova, S. N. Novikova, L. I. Lihacheva, O. M. Suhorada, T. A. Ruban, J. A. Kozel, D. M. Irodov, V. A. Kordium
Non-viral gene delivery of human apoA1 into mammalian cells in vitro and in vivo
Some approaches to the gene therapy of atherosclerosis by non-viral transfer of human apoA1 gene, encoding the main protein component of antiatherogenic lipoproteins, into liver cells are being studied. The plasmid vector for the expression of the human genomic apoA1 gene in mammalian cells has been constructed. The construction contains bacterial plasmid sequences for its replication in Escherihia coli and expression cassette for human apoA1 gene flanked by inverted terminal repeats of human adeno-associated virus. The cationic polymer polyethylenimine of 25 kD (PEI) has been used for gene delivery. It has been shown, that 50-70 % of cells express transgene after PEI-mediated in vitro transfection with analogical vector containing reporter green fluorescence protein gene. The experiments on animals have demonstrated, that the human protein ApoA1 is being tested in animal plasma during 15 days after apoA1 gene injection in the liver. PCR-analysis of total liver DNA of experimental animal has shown the presence of expected fragment of apoA1 gene in rat liver in 15 days after the gene delivery.