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利用融合脂质体将基因高效导入哺乳动物细胞。

Efficient gene transfer into mammalian cells using fusogenic liposome.

作者信息

Mizuguchi H, Nakagawa T, Nakanishi M, Imazu S, Nakagawa S, Mayumi T

机构信息

Faculty of Pharmaceutical Science, Osaka University, Japan.

出版信息

Biochem Biophys Res Commun. 1996 Jan 5;218(1):402-7. doi: 10.1006/bbrc.1996.0070.

Abstract

Fusogenic liposome (FL) based on Sendai virus constitutes a unique system that delivers the content efficiently into animal cells in vitro and in vivo. In this study we characterized unilamellar FL as a gene transfer vector in comparison with cationic lipid (CL)-DNA complex. FL transferred genes efficiently into cultured cells even when incubated for as little as 10 min, while CL-DNA complex required at least 30 min to reach the same level of gene expression. FL was also much less cytotoxic than CL-DNA complex under the conditions that resulted in the same level of gene expression. In addition, FL maintained 70% of the transfection activity even in the presence of 40% fetal calf serum (FCS), while CL-DNA complex almost completely lost their activity in the presence of 5% FCS. Furthermore, we found that FL could introduce and express luciferase gene into mouse ascites tumor cells in vivo, but CL-DNA complex could not even at higher concentrations of DNA. We conclude that unilamellar FL is a unique and efficient nonviral vector for gene transfer in vitro and in vivo.

摘要

基于仙台病毒的融合脂质体(FL)构成了一种独特的系统,可在体外和体内将内容物高效递送至动物细胞。在本研究中,我们将单层FL作为一种基因传递载体,并与阳离子脂质(CL)-DNA复合物进行了比较。即使仅孵育10分钟,FL也能将基因高效转移至培养细胞中,而CL-DNA复合物至少需要30分钟才能达到相同水平的基因表达。在导致相同基因表达水平的条件下,FL的细胞毒性也比CL-DNA复合物小得多。此外,即使在存在40%胎牛血清(FCS)的情况下,FL仍保持70%的转染活性,而CL-DNA复合物在存在5% FCS时几乎完全丧失活性。此外,我们发现FL可在体内将荧光素酶基因导入并表达于小鼠腹水肿瘤细胞中,但即使在更高的DNA浓度下,CL-DNA复合物也无法做到。我们得出结论,单层FL是一种独特且高效的非病毒载体,可用于体外和体内的基因传递。

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