Characters of dendritic poly(L-lysine) analogues with the terminal lysines replaced with arginines and histidines as gene carriers in vitro
Characters of dendritic poly(L-lysine) analogues with the terminal lysines replaced with arginines and histidines as gene carriers in vitro
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DOI：
10.1016/s0142-9612(03)00542-8
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发表时间：
2004-02-01
期刊：
影响因子：
14
通讯作者：
Aoyagi, H
中科院分区：
文献类型：
--
作者：
Okuda, T;Sugiyama, A;Aoyagi, H
非病毒基因传递系统的开发是实现治疗基因安全传递的重要关键，而没有病毒载体通常指出的副作用。我们已经表明，第6代（KG 6）的树突状聚（L-赖氨酸）显示出高转染效率到几个培养细胞具有低细胞毒性。在这里，为了研究取代末端阳离子基团对基因递送到细胞中的影响，我们合成了KGR 6和KGH 6，其中末端氨基酸分别被精氨酸和组氨酸取代。DNA结合分析表明，KGR 6与质粒DNA的结合能力与KG 6相当，而KGH 6的结合能力较低。KGR 6在几种培养细胞中的转染效率比KG 6高3至12倍。相比之下，KGH 6没有显示转染效率。然而，一旦KGH 6与DNA在酸性条件（pH 5.0）下混合，就形成DNA复合物，并且与KG 6介导的转染相比，它们显示出高转染效率。在酸性条件下形成的KGH 6的DNA复合物为1-2 μ m和球形，并且在中性条件下相对稳定。配合物的大小和球形与KG 6相同。KGH 6的独特性质将为我们构建体内外功能性基因转染系统提供基础和有价值的工具之一。(C)2003爱思唯尔有限公司。保留所有权利。
The development of a non-viral gene delivery system into cells is an important key to realize the safe delivery of therapeutic genes without the side effects often pointed out for viral vectors. We have shown that dendritic poly(L-lysine) of the 6th generation (KG6) shows high transfection efficiency into several cultivated cells with low cytotoxicity. Here, to investigate the effect of substituting terminal cationic groups on the gene delivery into cells, we synthesized KGR6 and KGH6, in which terminal amino acids were replaced by arginines and histidines, respectively. DNA-binding analysis showed that KGR6 could bind to the plasmid DNA as strongly as KG6, whereas KGH6 showed decreased binding ability. KGR6 showed 3- to 12-fold higher transfection efficiency into several cultivated cells than KG6. In contrast, KGH6 showed no transfection efficiency. However, once KGH6 was mixed with the DNA under acidic conditions (pH 5.0), DNA-complexes were formed and they showed high transfection efficiency compared to that in KG6-mediated transfection. DNA-complexes of KGH6 formed under acidic conditions were 1-2 mum and spherical, and relatively stable under neutral conditions. The size and spherical shape of the complexes were the same as those of KG6. The unique character of KGH6 will be one of the basic and valuable tools which will enable us to construct a functional gene transfection system in vitro and in vivo. (C) 2003 Elsevier Ltd. All rights reserved.