202. Effect of Modification with Lipophilic Fragments and Inclusion of Endosomolytic Peptide on Transfection Efficiency Mediated by Lysine Dendrimers

Abstract

Development of effective non-viral DNA carriers is considered to be a perspective approach for DNA delivery into eucaryotic cells with gene therapy aims. Our research was devoted to group of asymmetric lysine dendrimers. We studied fifth generation dendrimer D5 and its analog D5C10 which was modified by capric acid residues in external sphere. All vehicles were investigated for their capacity to bind DNA (using gel-retardation and ethidium bromide exclusion assay) and to provide protection plasmid DNA from nuclease degradation (using DNAse I protection test). Transfectional capacity of dendrimers were tested according to the efficiency of beta- galactosidase gene expression in HeLa cells. Modification of dendrimers by capric residues resulted in increase of ability to bind and to protect DNA from enzymatic degradation. Besides, D5C10 provides 10-fold more effective DNA delivery into cells, compared to D5. Addition of endosomolytic peptide JTS-1 to DNA/dendrimer complexes resulted in significant increase of gene delivery with dendrimers, especially with D5C10. The transfection effectiveness for DNA/D5C10/JTS-1 reached 16%. Thus we demonstrate that D5 and D5C10 carriers are perspective vehicles for DNA delivery into cells and can be basis for the development of more effective modular non-viral systems.