Human-derived NLS enhance the gene transfer efficiency of chitosan

Nuclear import is considered one of the major limitations for non-viral gene delivery systems and the incorporation of nuclear localization signals (NLS) that mediate nuclear intake can be used as a strategy to enhance internalization of exogenous DNA. In this work, human-derived endogenous NLS peptides based on Insulin Growth Factor Binding Proteins (IGFBP), namely IGFBP-3 and IGFBP-5, were tested for their ability to improve nuclear translocation of genetic material by non-viral vectors. Several strategies were tested to determine their effect on chitosan mediated transfection efficiency: co-administration with polyplexes, co-complexation at the time of polyplex formation, and covalent ligation to chitosan. Our results show that co-complexation and covalent ligation of human-derived NLS peptides to chitosan polyplexes yields a 2-fold increase in transfection efficiency. These results indicate that the integration of IGFBP-NLS peptides into polyplexes has potential as a strategy to enhance the efficiency of non-viral vectors.