Self-assembly of a multifunctional lipid with core-shell dendrimer DNA nanoparticles enhanced efficient gene delivery at low charge ratios into RPE cells

摘要

Gene therapy is a promising approach to treat genetic diseases. Development of gene delivery systems with high efficiency and low toxicity is a high priority in clinical application of gene therapy. In this work, we designed and evaluated a hybrid nonviral gene delivery system composed of a multifunctional lipid ECO and a core-shell dendrimer G4 nanoglobule for efficient intracellular gene delivery at low charge ratios (N/P). The dendrimeric nanoglobule was designed to effectively condense the gene cargo and the cationic lipid was used to enhance cellular uptake. As expected, this hybrid system formed stable nanoparticles at low N/P ratios (overall N/P = 6) and had low cytotoxicity. The hybrid nanoparticles induced higher GFP (green fluorescent protein) expression in ARPE-19 cells in the culture medium containing 10% serum than in serum-free medium at low N/P ratios. The nanoparticles also mediated significant reporter gene GFP expression in the retina ex-vivo from wild type C57 mice and in vivo with BALB/c mice. Together, these results demonstrate that the rationally designed G4 nanoglobule/pDNA/ECO nanoparticles are a promising delivery system for in vitro and in vivo gene delivery at low charge ratios.