Effect of lipid compositions on gene transfer into 293 cells using Sendai F/HN-virosomes.
Effect of lipid compositions on gene transfer into 293 cells using Sendai F/HN-virosomes.
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使用仙台 F/HN 病毒体，脂质成分对基因转移至 293 细胞的影响。
DOI：
10.5483/bmbrep.2002.35.5.459
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Y. Park
中科院分区：
文献类型：
--
作者：
Hong;Y. Park
融合仙台病毒包膜蛋白的融合脂质体，即所谓的仙台病毒体，已被开发用于动物细胞的体外和体内遗传修饰。在这项研究中，配制了几种具有不同脂质成分的不同病毒体，并比较了它们的体外基因转移效率。通过将仙台包膜、融合蛋白（F）和血凝素-神经氨酸酶（HN）蛋白定量重建到脂质体囊泡中来制备病毒体。在 293 个转化的人肾细胞中测试了含有荧光素酶报告基因的病毒体。用人工仙台病毒包膜（ASVE-病毒体）或磷脂酰丝氨酸（PS-病毒体）制备的F/HN-病毒体的基因转移效率比用1,2-二油酰-3-三甲基铵-丙烷（DOTAP）制备的阳离子脂质体高8或6倍。用磷脂酸（PA-病毒体）代替PS制备的F/HN病毒体在基因转移方面的效率低于ASVE-或PS-病毒体。此外，ASVE 和 PS 病毒体的基因转移能力在 Ca2+ 浓度为 510 mM 时达到最大。这些结果表明，掺入的脂质成分显着影响仙台 F/HN 病毒体介导的体外基因转移。
Fusogenic liposomes that incorporate Sendai virus envelope proteins, so-called Sendai virosomes, have been developed for in vitro and in vivo genetic modification of animal cells. In this study, several different virosomes of varying lipid compositions were formulated and their in vitro gene-transfer efficiencies compared. The virosomes were prepared by quantitative reconstitution of the Sendai envelope, fusion (F) and hemagglutinin-neuraminidase (HN) proteins into liposomal vesicles. Virosomes that contained luciferase reporter genes were tested in 293 transformed human kidney cells. F/HN-virosomes that were prepared with an artificial Sendai viral envelope (ASVE-virosomes) or phosphatidylserine (PS-virosomes) exhibited an 8- or 6-fold higher gene-transfer efficiency than cationic liposomes that were made with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). F/HNvirosomes that were prepared with phosphatidic acid (PA-virosomes) instead of PS were less efficient in gene transfer than either ASVE- or PS-virosomes. In addition, the gene-transfer capability of ASVE- and PS-virosomes was maximal at a Ca2+ concentration of 510 mM. These results suggest that the incorporated lipid components significantly affect the in vitro gene transfer that is mediated by Sendai F/HN-virosomes.