Department of Animal and Veterinary Science, University of Idaho, Moscow, ID, USA.
Department of Animal and Veterinary Science, University of Idaho, Moscow, ID, USA.
Nanomedicine. 2014 Feb;10(2):329-37. doi: 10.1016/j.nano.2013.07.015. Epub 2013 Aug 6.
Gold nanoparticles (AuNP) were conjugated with cysteine terminated KDEL (Lys-Asp-Glu-Leu) peptide and siRNA directed against NADPH Oxidase 4 (Nox4). Fluorescence microscopy analysis provided evidence of cytocellular retrograde transport pathways and sub-cellular colocalization of AuNP nanoconstructs in both undifferentiated C2C12 myoblasts and differentiated C2C12 myotubes. The cellular trafficking of AuNP nanoconstructs in undifferentiated myoblasts suggests stable and efficient transfection of siRNA as demonstrated by colocalization of AuNP-delivered KDEL and siRNA. The cellular uptake of AuNP nanoconstructs was more efficient than Lipofectamine mediated transfection in differentiated myotubes (P<0.05) compared to undifferentiated myoblasts, suggesting that AuNP nanoconstructs provide an efficient platform for siRNA delivery to differentiated myotubes. The localization of these nanoconstructs in undifferentiated myoblasts suggests that most of the siRNA was localized in the endoplasmic reticulum (ER) with a minimal distribution in the Golgi bodies suggesting that the ER is a primary localization site for AuNP-KDEL mediated delivery of nanoconstructs.
This team of investigators demonstrate that a delivery system composed of gold nanoparticle and KDEL based siRNA is superior to lipofectamine in delivering siRNA in differentiated myoblasts, paving the way to gene silencing methods in these cells for future therapeutic exploitation.
金纳米粒子（AuNP）与半胱氨酸末端 KDEL（Lys-Asp-Glu-Leu）肽和针对 NADPH 氧化酶 4（Nox4）的 siRNA 缀合。荧光显微镜分析提供了证据，表明 AuNP 纳米结构在未分化的 C2C12 成肌细胞和分化的 C2C12 肌管中存在细胞内逆行运输途径和亚细胞共定位。AuNP 纳米结构在未分化的成肌细胞中的细胞内转运表明 siRNA 的稳定和高效转染，这表现为 AuNP 递呈的 KDEL 和 siRNA 的共定位。与未分化的成肌细胞相比，AuNP 纳米结构在分化的肌管中的细胞摄取效率高于脂质体介导的转染（P<0.05），这表明 AuNP 纳米结构为分化的肌管中 siRNA 的递呈提供了高效平台。这些纳米结构在未分化的成肌细胞中的定位表明，大多数 siRNA 定位于内质网（ER）中，高尔基体中的分布最小，这表明 ER 是 AuNP-KDEL 介导的纳米结构递呈的主要定位部位。
该研究小组的研究人员证明，由金纳米粒子和基于 KDEL 的 siRNA 组成的递药系统在递呈 siRNA 方面优于脂质体，为这些细胞中的基因沉默方法铺平了道路，以便未来进行治疗性开发。
