Cellular fate of a modular DNA delivery system mediated by silica nanoparticles
Cellular fate of a modular DNA delivery system mediated by silica nanoparticles
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DOI：
10.1021/bp049648w
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发表时间：
2005-03-01
影响因子：
2.9
通讯作者：
Saltzman, WM
中科院分区：
文献类型：
--
作者：
Gemeinhart, RA;Luo, D;Saltzman, WM
有效的分子药物，包括基因治疗剂、RNA治疗剂和DNA疫苗的开发，依赖于将DNA或RNA转移到细胞中的有效手段。潜在的问题，包括毒性和免疫原性，周围的病毒方法的DNA交付有必要使用非病毒，合成载体。为了更好地设计合成载体或转染试剂，病毒的模块化设计激发了DNA和RNA递送的模块化方法。每个模块化组件都可以被设计成绕过许多障碍中的每一个。模块化方法将允许为特定应用修改单个组件。通过利用致密的二氧化硅纳米颗粒形成三元复合物，DNA-转染试剂复合物的转染效率通过在细胞表面浓缩DNA而增加约10倍。二氧化硅纳米颗粒的表面改性允许测定细胞摄取机制，转染效率只有微小的改变。纳米颗粒通过核内体-溶酶体途径内化，然后在核周积聚。修饰机制证实模块化系统的表面修饰可以允许特定部分并入模块化系统中，而不会显著改变修饰效率。通过显示基于细胞水平的DNA浓度的模块化系统可用于增加转染效率，我们已经显示了系统的进一步修饰可以更好地靶向DNA递送并克服DNA表达的其他障碍。
Development of efficient molecular medicines, including gene therapeutics, RNA therapeutics, and DNA vaccines, depends on efficient means of transfer of DNA or RNA into the cell. Potential problems, including toxicity and immunogenicity, surrounding viral methods of DNA delivery have necessitated the use of nonviral, synthetic carriers. To better design synthetic carriers, or transfection reagents, the modular design of viruses has inspired a modular approach to DNA and RNA delivery. Each modular component can be designed to circumvent each of the many barriers. The modular approach will allow modification of individual components for a specific application. By utilizing a dense silica nanoparticle to form a ternary complex, transfection efficiency of a DNA-transfection reagent complex was increased by a factor of approximately 10 by concentrating the DNA at the surface of cells. Surface modification of the silica nanoparticles allowed determination of the cellular uptake mechanism with only minor alteration of transfection efficiency. Nanoparticles are internalized by an endosome-lysosomal route followed by perinuclear accumulation. The modification mechanism confirms that surface modification of the modular system can allow specific moieties to be incorporated into the modular system without significant alteration of the transfbetion efficiency. By showing that the modular system based upon concentration of DNA at the level of the cell can be used to increase transfection efficiency, we have shown that further modification of the system may better target DNA delivery and overcome other barriers of DNA expression.