Tiscornia Gustavo, Singer Oded, Verma Inder M
The Salk Institute for Biological Studies, Laboratory of Genetics, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
Nat Protoc. 2006;1(1):241-5. doi: 10.1038/nprot.2006.37.
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.
慢病毒载体作为基因传递的工具，具有独特的多功能性和稳健性。它们可以转导多种细胞类型，并在分裂细胞和有丝分裂后细胞中整合到宿主基因组中，从而在体外和体内实现转基因的长期表达。本方案描述了慢病毒载体如何生产、纯化和滴定。高滴度悬浮液通常可以相对轻松地制备：在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（10⁶病毒颗粒/毫升）的未纯化制剂；高滴度（10⁹病毒颗粒/毫升）的纯化制剂则还需要2天时间。
