Effect of polyethyleneglycol spacer on the binding properties of nuclear localization signal-modified liposomes to isolated nucleus.
Effect of polyethyleneglycol spacer on the binding properties of nuclear localization signal-modified liposomes to isolated nucleus.
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聚乙二醇间隔基对核定位信号修饰脂质体与离体细胞核结合特性的影响。
DOI：
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发表时间：
2009
影响因子：
2
通讯作者：
H. Harashima
中科院分区：
文献类型：
--
作者：
Dai Kurihara;H. Akita;A. Kudo;T. Masuda;S. Futaki;H. Harashima
核递送过程是成功基因递送的关键障碍，特别是在非分裂细胞中。我们之前提出了一种用于质粒DNA（pDNA）核递送的新策略，其中pDNA被包裹在已用核靶向信号修饰的脂质双层中，包括源自SV 40（NLS）或糖单元的核定位信号。在本研究中，我们报告了脂质体修饰的NLS的拓扑结构对其与分离细胞核结合能力的影响的研究。通过掺入硬脂酰化NLS（STR-NSL）或通过用聚乙二醇（PEG）间隔基修饰（NLS-PEG-Lip）将NLS直接连接至脂质体（NLS-Lip）。使用NLS未修饰的脂质体（PEG-Lip）作为对照。用7-硝基苯-2-氧杂-1，3-二唑（NBD）标记后，将脂质体与来自JAWS II细胞的细胞匀浆一起孵育，然后通过离心分离核级分。PEG-Lip制剂显示与细胞核的结合可忽略不计。相反，NLS-Lips与细胞核的结合以STR-NLS密度依赖性方式逐渐增加。有趣的是，NLS-PEG-Lips与细胞核的结合即使在低密度下也是高度有效的，这表明PEG间隔物的存在是改善NLS修饰的脂质体与细胞核的结合活性的重要因素。这些信息将有助于核靶向载体的设计。
The nuclear delivery process is a crucial barrier to successful gene delivery, especially in non-dividing cells. We previously proposed a novel strategy for the nuclear delivery of plasmid DNA (pDNA), in which the pDNA is encapsulated in lipid bilayers that had been modified with nucleus-targeting signals, including nuclear localizing signals derived from SV40 (NLS) or sugar units. In the present study, we report on an investigation of the effect of the topology of the liposome-modified NLS on its ability to bind to the isolated nucleus. NLS was directly attached to a liposome (NLS-Lip) by incorporating stearylated NLS (STR-NSL), or by modification with a polyethyleneglycol (PEG) spacer (NLS-PEG-Lip). NLS-unmodified liposomes (PEG-Lip) were used as a control. The liposomes, after labeling with 7-nitrobenz-2-oxa-1,3-diazole (NBD), were incubated with a cell homogenate derived from JAWS II cells, followed by isolation of the nuclear fraction by centrifugation. The PEG-Lip preparation showed negligible binding to the nucleus. In contrast, the binding of NLS-Lips to the nucleus gradually increased in a STR-NLS density-dependent manner. Interestingly, the binding of NLS-PEG-Lips to the nucleus is highly effective even at low density, suggesting that the presence of the PEG spacer is an important factor in improving the binding activity of NLS-modified liposomes to the nucleus. This information will be useful for the design of nucleus-targeting carriers.
影响因子：
2.9
作者：
Wyman, TB;Nicol, F;Szoka, FC
通讯作者：
Szoka, FC