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Production and purification of lentiviral vectors

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Abstract

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 (106 viral particles/ml) unpurified preparation can be obtained 3 d after transfecting cells with lentiviral vector and packaging plasmids; a high-titer (109 viral particles/ml) purified preparation requires 2 more days.

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Figure 1: Schematic representation of the third generation lentiviral system.

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Authors and Affiliations

  1. The Salk Institute for Biological Studies, Laboratory of Genetics, 10010 North Torrey Pines Road, La Jolla, 92037, California, USA

    Gustavo Tiscornia, Oded Singer & Inder M Verma

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  1. Gustavo Tiscornia
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  2. Oded Singer
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  3. Inder M Verma
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Correspondence to Inder M Verma.

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The authors declare no competing financial interests.

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Tiscornia, G., Singer, O. & Verma, I. Production and purification of lentiviral vectors. Nat Protoc 1, 241–245 (2006). https://doi.org/10.1038/nprot.2006.37

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