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Generation of transgenic birds with replication-deficient lentiviruses

Nature Protocols volume 1, pages 1406–1411 (2006)Cite this article

Abstract

Birds are of great interest as an animal model in biological research and for commercial applications as a bioreactor. Effective methods for manipulating the avian genome would accelerate progress in fields such as developmental biology and behavioral neurobiology, which traditionally have relied on birds as model systems for biological research. Here we describe a simple and effective protocol for producing transgenic birds using lentiviral vectors that can be used to achieve tissue-specific transgene expression at high levels. The time allotted for the procedure depends upon the species of bird; adult transgenic quails can be generated in approximately 5 months.

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Figure 1: Diagram of the egg injection process, Steps 7–9.
Figure 2: GFP fluorescence in E6 embryos from two strains of transgenic quails observed by fluorescence microscopy.
Figure 3: Southern blot analysis of genomic DNA from F1 and F2 transgenic quails.

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References

  1. Salter, D.W., Smith, E.J., Hughes, S.H., Wright, S.E. & Crittenden, L.B. Transgenic chickens: insertion of retroviral genes into the chicken germ line. Virology 157, 236–240 (1987).

    Article  CAS  Google Scholar 

  2. Jaenisch, R. Germ line integration and Mendelian transmission of the exogenous Moloney leukemia virus. Proc. Natl. Acad. Sci. USA 73, 1260–1264 (1976).

    Article  CAS  Google Scholar 

  3. Bosselman, R.A. et al. Germline transmission of exogenous genes in the chicken. Science 243, 533–535 (1989).

    Article  CAS  Google Scholar 

  4. Mizuarai, S. et al. Production of transgenic quails with high frequency of germ-line transmission using VSV-G pseudotyped retroviral vector. Biochem. Biophys. Res. Commun. 286, 456–463 (2001).

    Article  CAS  Google Scholar 

  5. Ivarie, R. Avian transgenesis: progress towards the promise. Trends Biotechnol. 21, 14–19 (2003).

    Article  CAS  Google Scholar 

  6. van de Lavoir, M.C. et al. Germline transmission of genetically modified primordial germ cells. Nature 441, 766–769 (2006).

    Article  CAS  Google Scholar 

  7. McGrew, M.J. et al. Efficient production of germline transgenic chickens using lentiviral vectors. EMBO Rep. 5, 728–733 (2004).

    Article  CAS  Google Scholar 

  8. Chapman, S.C. et al. Ubiquitous GFP expression in transgenic chickens using a lentiviral vector. Development 132, 935–940 (2005).

    Article  CAS  Google Scholar 

  9. Scott, B.B. & Lois, C. Generation of tissue-specific transgenic birds with lentiviral vectors. Proc. Natl. Acad. Sci. USA 102, 16443–16447 (2005).

    Article  CAS  Google Scholar 

  10. Lois, C., Hong, E.J., Pease, S., Brown, E.J. & Baltimore, D. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295, 868–872 (2002).

    Article  CAS  Google Scholar 

  11. Hofmann, A. et al. Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep. 4, 1054–1060 (2003).

    Article  CAS  Google Scholar 

  12. Hofmann, A. et al. Generation of transgenic cattle by lentiviral gene transfer into oocytes. Biol. Reprod. 71, 405–409 (2004).

    Article  CAS  Google Scholar 

  13. Lois, C. Generation of transgenic animals using lentiviral vectors. In Mammalian and avian transgenesis: new approaches (eds., Pease, S. & Lois, C.) 1–22 (Springer, Berlin, 2005).

    Google Scholar 

  14. Speksnijder, G. & Ivarie, R. A modified method of shell windowing for producing somatic or germline chimeras in fertilized chicken eggs. Poult. Sci. 79, 1430–1433 (2000).

    Article  CAS  Google Scholar 

  15. Andacht, T., Hu, W. & Ivarie, R. Rapid and improved method for windowing eggs accessing the stage X chicken embryo. Mol. Reprod. Dev. 69, 31–34 (2004).

    Article  CAS  Google Scholar 

  16. Sambrook, J. & Russel, D.W. Molecular cloning: a laboratory manual 3rd edn. (Cold Spring Harbor Press, Cold Spring Harbor, 2001).

    Google Scholar 

  17. Bellairs, R. & Osmond, M. Atlas of chick development 2nd edn. (Elsevier Academic Press, London, 2005).

    Google Scholar 

Download references

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Authors and Affiliations

  1. Picower Institute for Learning and Memory MIT, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Benjamin B Scott & Carlos Lois

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  1. Benjamin B Scott
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  2. Carlos Lois
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Correspondence to Carlos Lois.

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The authors declare no competing financial interests.

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Scott, B., Lois, C. Generation of transgenic birds with replication-deficient lentiviruses. Nat Protoc 1, 1406–1411 (2006). https://doi.org/10.1038/nprot.2006.187

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