Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan.
Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu 30013, Taiwan.
Biomaterials. 2014 Sep;35(28):8261-72. doi: 10.1016/j.biomaterials.2014.06.016. Epub 2014 Jun 25.
Gene therapy provides a new hope for previously "incurable" diseases. Low gene transfection efficiency, however, is the bottle-neck to the success of gene therapy. It is very challenging to develop non-viral nanocarriers to achieve ultra-high gene transfection efficiencies. Herein, we report a novel design of "tight binding-but-detachable" lipid-nanoparticle composite to achieve ultrahigh gene transfection efficiencies of 60∼82%, approaching the best value (∼90%) obtained using viral vectors. We show that Fe@CNPs nanoparticles coated with LP-2000 lipid molecules can be used as gene carriers to achieve ultra-high (60-80%) gene transfection efficiencies in HeLa, U-87MG, and TRAMP-C1 cells. In contrast, Fe@CNPs having surface-covalently bound N,N,N-trimethyl-N-2-methacryloxyethyl ammonium chloride (TMAEA) oligomers can only achieve low (23-28%) gene transfection efficiencies. Similarly ultrahigh gene transfection/expression was also observed in zebrafish model using lipid-coated Fe@CNPs as gene carriers. Evidences for tight binding and detachability of DNA from lipid-nanoparticle nanocarriers will be presented.
基因治疗为以前的“不治之症”提供了新的希望。然而，低基因转染效率是基因治疗成功的瓶颈。开发非病毒纳米载体以实现超高基因转染效率极具挑战性。在此，我们报告了一种新型“紧密结合但可分离”的脂质-纳米颗粒复合材料的设计，以实现超高的基因转染效率（60∼82%），接近使用病毒载体获得的最佳值（∼90%）。我们表明，涂有 LP-2000 脂质分子的 Fe@CNPs 纳米颗粒可用作基因载体，在 HeLa、U-87MG 和 TRAMP-C1 细胞中实现超高（60-80%）的基因转染效率。相比之下，表面共价键合 N,N,N-三甲基-N-2-甲基丙烯酰氧乙基氯化铵（TMAEA）低聚物的 Fe@CNPs 只能实现低（23-28%）的基因转染效率。同样，使用脂质涂覆的 Fe@CNPs 作为基因载体，在斑马鱼模型中也观察到超高的基因转染/表达。我们将展示 DNA 与脂质-纳米颗粒纳米载体紧密结合和可分离的证据。
