The effect of nocodazole on the transfection efficiency of lipid-bilayer coated gold nanoparticles
The effect of nocodazole on the transfection efficiency of lipid-bilayer coated gold nanoparticles
复制标题
诺考达唑对脂质双层包被的金纳米粒子转染效率的影响。
DOI：
10.1016/j.biomaterials.2008.11.037
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发表时间：
2009-03-01
期刊：
影响因子：
14
通讯作者：
Wang, Erkang
中科院分区：
文献类型：
--
作者：
Li, Dan;Li, Peicai;Wang, Erkang
在基因治疗应用中，非病毒载体比病毒系统更安全。但由于其疗效有限，一直阻碍着其在临床的广泛应用。除了寻找新的基因非病毒载体外，许多研究者还致力于寻找新的物质来提高现有载体的转染效率。在这项工作中，我们发现了一种转染增强剂nocodazole (NCZ)，用于二甲二十二烷基铵（DODAB），一种阳离子脂质双分子层包覆的金纳米颗粒（AuNPs）介导的基因传递。流式细胞术检测发现NCZ对HEK 293T细胞的转染增强作用为3倍。荧光素酶实验进一步证实了这一结果，转染48 h后NCZ的转染效率提高了5倍以上。电感耦合等离子体质谱（ICP-MS）和FCM分析结果表明，NCZ对DODAB-AuNPs/DNA复合物的内在化没有影响。通过透射电镜观察复合物的运输情况，发现复合物向溶酶体的运输中断是转染增强的主要原因。因此。NCZ可以在DODAB-AuNPs介导的转染系统中作为转染增强子。这项工作也为提高脂质介导转染的效率提供了见解：在转染前修饰金纳米颗粒上的脂质并用NCZ预处理细胞。(c) 2008 Elsevier Ltd.版权所有。
Nonviral vectors are safer than viral systems for gene therapy applications. However, the limited efficacy always prevents their being widely used in clinical practice. Aside from searching new gene nonviral vectors, many researchers focus on finding out new substances to improve the transfection efficiency of existent vectors. In this work, we found a transfection enhancer, nocodazole (NCZ), for dimethyldioctadecylammonium (DODAB, a cationic lipid) bilayer coated gold nanoparticles (AuNPs) mediated gene delivery. It was found that NCZ produces 3-fold transfection enhancement to HEK 293T cells assessed by flow cytometry (FCM). The result was further confirmed by luciferase assay, in which NCZ induced more than 5 times improvement in transfection efficiency after 48 h of transfection. The results from the inductively coupled plasma mass spectrometry (ICP-MS) and FCM showed that NCZ did not affect the internalization of DODAB-AuNPs/DNA complexes. The trafficking of the complexes by transmission electron microscopy (TEM) indicated that the interrupted transportation of the complexes to the lysosomes contributed greatly to the transfection enhancement. Therefore. NCZ can be used as a transfection enhancer in DODAB-AuNPs mediated transfection system. This work also gave an insight to improving the efficiency of lipid-mediated transfection: modifying lipid on gold nanoparticles and pretreating cells by NCZ before the transfection. (c) 2008 Elsevier Ltd. All rights reserved.