Nuclear location signal peptide-modified poly (ethyleneimine)/DNA complexes: An efficient gene delivery vector in vitro and in vivo

The low transfection efficiency of nonviral gene delivery systems limits their applications. In this study, we demonstrated a simple method to modify poly(ethyleneimine)/DNA complexes with a nuclear location signal peptide via bis(succinimidyl) penta(ethylene glycol) coupling. The amount of grafted nuclear location signal peptide was controlled within a range of 0-9 mu g for poly(ethyleneimine)/DNA complexes containing 10 mu g DNA and 100 mu g poly(ethyleneimine) by adjusting the grafting agent and peptide feeds. The particle size and surface zeta-potential of the complexes were largely retained after nuclear location signal immobilization. Based on the results of the flow cytometry measurements, the nuclear location signal-modified poly(ethyleneimine)/DNA complexes were internalized into at bone marrow stem cells at a significantly faster rate and a higher amount than the unmodified complexes. In vitro transfection experiments, using plasmid DNA encoding bone morphogenetic protein 2, indicated that the nuclear location signal peptide-modified poly(ethyleneimine)/DNA complexes have significantly higher gene transfection ability toward bone marrow stem cells than unmodified complexes. The porous collagen scaffolds loaded with nuclear location signal-modified poly(ethyleneimine)/plasmid DNA encoding bone morphogenetic protein 2 complexes successfully transfected tissue cells and induced the human bone morphogenetic protein 2 expression in a rat. The modification of the poly(ethyleneimine)/DNA complexes with nuclear location signal peptide was effective in enhancing gene transfection of complexes in vitro and in vivo, thus indicating potential applications for bioactive scaffolds with enhanced tissue regeneration performance.