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Rapid generation of knockdown transgenic mice by silencing lentiviral vectors

Nature Protocols volume 1, pages 286–292 (2006)Cite this article

Abstract

Lentiviral vectors are potent gene delivery vehicles that enable stable expression of transgenes in both dividing and postmitotic cells, including preimplantation embryos. We have developed lentiviral vectors carrying silencing cassettes consisting of an RNA polymerase III promoter expressing short hairpin RNAs. Transgenic mice can be generated rapidly by transduction of early embryos with lentiviral silencing vectors, resulting in mice with downregulated target genes. We describe two alternative early embryo transduction protocols (removal of zona pellucida and subzonal microinjection). These methodologies offer the possibility of large-scale generation of knockdown transgenic mice for functional genomic studies and enable the production of transgenic mice in 7 weeks.

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Figure 1: Manipulation of mouse embryos, adapted from ref. 9.
Figure 2: Subzonal injections of lentiviral particles.
Figure 3: Analysis of efficiency of transgenesis in a group of littermate F1 mice born from a cross between two founder mice.

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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

    Oded Singer, Gustavo Tiscornia, Masahito Ikawa & Inder M Verma

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  1. Oded Singer
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  2. Gustavo Tiscornia
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  3. Masahito Ikawa
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  4. 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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Singer, O., Tiscornia, G., Ikawa, M. et al. Rapid generation of knockdown transgenic mice by silencing lentiviral vectors. Nat Protoc 1, 286–292 (2006). https://doi.org/10.1038/nprot.2006.44

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