Production and purification of lentiviral vectors
Production and purification of lentiviral vectors
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DOI：
10.1038/nprot.2006.37
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Verma, Inder M.
中科院分区：
文献类型：
--
作者：
Tiscornia, Gustavo;Singer, Oded;Verma, Inder M.
慢病毒载体提供了独特的多功能性和健壮性作为载体的基因传递。它们可以转导广泛的细胞类型，并在分裂和有丝分裂后的细胞中整合到宿主基因组中，从而在体外和体内长期表达转基因。本方案描述了慢病毒载体如何产生、纯化和滴定。高滴度的悬浮液可以相对容易地常规制备：用慢病毒载体和包装质粒转染细胞3天后，可以获得低滴度（10(6)个病毒颗粒/ ml）的未纯化制剂；高滴度（10(9)个病毒颗粒/ ml）的纯化制剂需要2天以上。
Lentiviral vectors offer unique versatility and robustness as vehicles for gene delivery. They can transduce a wide range of cell types and integrate into the host genome in both dividing and post- mitotic cells, resulting in long- term expression of the transgene both in vitro and in vivo. This protocol describes how lentiviral vectors can be produced, purified and titrated. High titer suspensions can be routinely prepared with relative ease: a low- titer ( 10(6) viral particles/ ml) unpurified preparation can be obtained 3 d after transfecting cells with lentiviral vector and packaging plasmids; a high- titer ( 10(9) viral particles/ ml) purified preparation requires 2 more days.