NLS Peptide Improves the Efficiency of pDNA Delivery into Eukaryotic Cells Mediated by Cationic Liposomes

Abstract— Objective: The main limitation of DNA use in the therapy of genetic and acquired diseases is the development of its effective delivery systems. The aim of this work is to evaluate the effect of the NLS peptide (CKRPAATKKAGQAKKKK) on the efficiency of pDNA delivery by conventional and multifunctional cationic liposomes into eukaryotic cells. Methods: The effect of the NLS peptide on the efficiency of pDNA binding in the presence and absence of cationic liposomes was studied using gel electrophoresis and dynamic laser light scattering. Complexes of cationic liposomes were formed with pDNA in the presence or absence of the NLS peptide at different component ratios (N/P). The transfection efficiency of the complexes in HEK 293 and KB-3-1 cells was studied using flow cytometry. Results and Discussion: In the case of using NLS for targeted delivery of pDNA (at low N/P ratios) into KB-3-1 cells, the most optimal systems are multifunctional cationic liposomes F2P2 containing 2 mol % folate and 2 mol % PEG-lipid Р800. At higher N/P ratios in the case of accumulation of liposome complexes with pDNA/NLS, the most optimal systems were conventional cationic liposomes L for HEK 293 cells and PEGylated liposomes P4 containing 4% PEG-lipid P800 for KB-3-1 cells. Conclusions: The obtained data can be used to solve the problem of efficient delivery of pDNA into eukaryotic cells and increase the efficiency of protein expression by 1.5–2 times.