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日本血凝病毒(HVJ)包膜载体作为一种通用的基因递送系统。

Hemagglutinating virus of Japan (HVJ) envelope vector as a versatile gene delivery system.

作者信息

Kaneda Yasufumi, Nakajima Toshihiro, Nishikawa Tomoyuki, Yamamoto Seiji, Ikegami Hiroyuki, Suzuki Naho, Nakamura Hitomi, Morishita Ryuichi, Kotani Hitoshi

机构信息

Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, Suita, Osaka, 565-0871, Japan.

出版信息

Mol Ther. 2002 Aug;6(2):219-26. doi: 10.1006/mthe.2002.0647.

Abstract

We have developed a simple method for converting the lipid envelope of an inactivated virus to a gene transfer vector. Hemagglutinating virus of Japan (HVJ; Sendai virus) envelope vector was constructed by incorporating plasmid DNA into inactivated HVJ particles. This HVJ envelope vector introduced plasmid DNA efficiently and rapidly into various cell lines, including cancer cells and several types of primary cell culture. Efficiency of gene transfer was greatly enhanced by protamine sulfate and centrifugation. Fluorescein isothiocyanate-labeled oligodeoxynucleotides (FITC-ODN) were also delivered to cells at > 95% efficiency. When HVJ envelope vector was injected into organs directly, reporter gene expression was observed in organs including liver, brain, skin, uterus, tumor masses, lung, and eye. When HVJ envelope vector containing luciferase gene was injected into mouse tail vein, luciferase gene expression was detected primarily in spleen. FITC-ODN were also delivered to spleen cells by intravenous injection of HVJ envelope. These results suggest that HVJ envelope vector will be useful for both ex vivo and in vivo gene therapy experiments.

摘要

我们已经开发出一种简单的方法,可将灭活病毒的脂质包膜转化为基因传递载体。通过将质粒DNA掺入灭活的日本血凝病毒(HVJ;仙台病毒)颗粒中构建了HVJ包膜载体。这种HVJ包膜载体能高效、快速地将质粒DNA导入各种细胞系,包括癌细胞和几种原代细胞培养物。硫酸鱼精蛋白和离心极大地提高了基因传递效率。异硫氰酸荧光素标记的寡脱氧核苷酸(FITC-ODN)也能以>95%的效率递送至细胞。当将HVJ包膜载体直接注射到器官中时,在包括肝脏、大脑、皮肤、子宫、肿瘤块、肺和眼睛等器官中观察到报告基因表达。当将含有荧光素酶基因的HVJ包膜载体注射到小鼠尾静脉中时,主要在脾脏中检测到荧光素酶基因表达。通过静脉注射HVJ包膜也能将FITC-ODN递送至脾细胞。这些结果表明,HVJ包膜载体将对离体和体内基因治疗实验都有用。

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