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通过将定量重组的仙台包膜蛋白载入脂质体进行基因转染。

Gene transfection by quantitatively reconstituted Sendai envelope proteins into liposomes.

作者信息

Kim Hong Sung, Park Yong Serk

机构信息

Department of Medical Technology, Yonsei University, Wonju 220-710, Republic of Korea.

出版信息

Cancer Gene Ther. 2002 Feb;9(2):173-7. doi: 10.1038/sj.cgt.7700421.

Abstract

Fusogenic liposomes (virosomes) consisting of Sendai virus envelope proteins have been utilized for in vitro and in vivo genetic modification of animal cells. In this study, the virosomes containing DNA were prepared by quantitative reconstitution of Sendai envelope proteins, fusion protein and hemagglutinin-neuramindase in liposomal vesicles. The Sendai virosomes more efficiently transferred genes into cultured 293 transformed kidney cells than 1,2-dioleoyl-3-(trimethylammonium) propane-based cationic liposomes. At 200:1 weight ratio of envelope protein and lipid, the virosomes exhibited the best efficiency of gene transfection into the cells. The Sendai virosomes required relatively a short period of incubation time and much less cytotoxic, compared to the cationic liposome/DNA complex. The transfection efficiency of the Sendai virosomes containing DNA was maintained 70% after a month. This type of Sendai virosomes is relatively convenient for preparation and storage, compared to fusogenic liposomes prepared by liposome-virus fusion. First of all, because the constituents are quantitatively formulated, this type of virosome formulation can provide further consistent transfection for gene therapy.

摘要

由仙台病毒包膜蛋白组成的融合脂质体(病毒体)已被用于动物细胞的体外和体内基因改造。在本研究中,通过在脂质体囊泡中定量重组仙台包膜蛋白、融合蛋白和血凝素 - 神经氨酸酶来制备含DNA的病毒体。与基于1,2 - 二油酰基 - 3 -(三甲基铵)丙烷的阳离子脂质体相比,仙台病毒体能更有效地将基因转移到培养的293转化肾细胞中。在包膜蛋白与脂质的重量比为200:1时,病毒体对细胞的基因转染效率最佳。与阳离子脂质体/DNA复合物相比,仙台病毒体所需的孵育时间相对较短,细胞毒性也小得多。含DNA的仙台病毒体在一个月后仍保持70%的转染效率。与通过脂质体 - 病毒融合制备的融合脂质体相比,这种类型的仙台病毒体在制备和储存方面相对方便。首先,由于成分是定量配制的,这种类型的病毒体制剂可为基因治疗提供更一致的转染效果。

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