Transposon-free insertions for insect genetic engineering (original) (raw)

Nature Biotechnology volume 24, pages 820–821 (2006)Cite this article

Abstract

Methods involving the release of transgenic insects in the field hold great promise for controlling vector-borne diseases and agricultural pests. Insect transformation depends on nonautonomous transposable elements as gene vectors. The resulting insertions are stable in the absence of suitable transposase, however, such absence cannot always be guaranteed. We describe a method for post-integration elimination of all transposon sequences in the pest insect Medfly, Ceratitis capitata. The resulting insertions lack transposon sequences and are therefore impervious to transposase activity.

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Figure 1: Strategy for post-integration removal of transposon ends from transposon-mediated insertions.

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References

  1. Li, X. et al. Insect Mol. Biol. 14, 17–30 (2005).
    Article Google Scholar
  2. Wimmer, E. Nat. Rev. Genet. 4, 225–232 (2003).
    Article CAS Google Scholar
  3. Atkinson, P., Pinkerton, A. & O'Brochta, D . Annu. Rev. Entomol. 46, 317–346 (2001).
    Article CAS Google Scholar
  4. Alphey, L. Insect Biochem. Mol. Biol. 32, 1243–1247 (2002).
    Article CAS Google Scholar
  5. Alphey, L. et al. Science 298, 119–121 (2002).
    Article CAS Google Scholar
  6. Olson, K.E., Alphey, L., Carlson, J. & James, A.A. in Bridging Laboratory and Field Research for Genetic Control of Disease Vectors, vol. 11 (eds. Knols, B.G.J. & Louis, C.) 210 (Springer, Heidelberg, 2006).
    Google Scholar
  7. Handler, A . Insect Biochem. Mol. Biol. 34, 121–130 (2004).
    Article CAS Google Scholar
  8. Hoy, M. in Insect transgenesis: methods and applications (eds. James, A. & Handler, A.) 335–367 (CRC Press, Boca Raton, 2000).
    Google Scholar
  9. Handler, A., Zimowska, G. & Horn, C. Nat. Biotechnol. 22, 1150–1154 (2004).
    Article CAS Google Scholar
  10. Li, X., Lobo, N., Bauser, C. & Fraser, M. Mol. Genet. Genomics 266, 190–198 (2001).
    Article CAS Google Scholar
  11. Berg, C. & Spradling, A. Genetics 127, 515–524 (1991).
    CAS PubMed PubMed Central Google Scholar
  12. Horn, C. & Handler, A. . Proc. Natl. Acad. Sci. USA 102, 12483–12488 (2005).
    Article CAS Google Scholar
  13. Oberstein, A., Pare, A., Kaplan, L. & Small, S. Nat. Methods 2, 583–585 (2005).
    Article CAS Google Scholar
  14. Nimmo, D., Alphey, L., Meredith, J. & Eggleston, P. Insect Mol. Biol. 15, 129–136 (2006).
    Article CAS Google Scholar
  15. Shagin, D.A. et al. Mol. Biol. Evol. 21, 841–850 (2004).
    Article CAS Google Scholar

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Acknowledgements

We thank Karen Clifton and staff at Oxitec for technical assistance, Nadia Pantic and Peng Gong for help at early stages of the project, Pedro Rendón for the Toliman Medfly strain and Tassos Mintzas, Sinead O'Connell and Derric Nimmo for advice and critical review of the manuscript. This work was funded in part by the UK Biotechnology and Biological Sciences Research Council.

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Author notes

  1. Tarig H Dafa'alla, George C Condon and Luke Alphey: These authors contributed equally to this work.

Authors and Affiliations

  1. Oxitec Limited, 71 Milton Park, Oxford, OX14 4RX, UK
    Tarig H Dafa'alla, George C Condon, Kirsty C Condon, Caroline E Phillips, Neil I Morrison, Li Jin, Guoliang Fu & Luke Alphey
  2. Department of Zoology, University of Oxford, South Parks Road, Oxford, OX1 3PS, UK
    Kirsty C Condon, Neil I Morrison, Matthew J Epton & Luke Alphey

Authors

  1. Tarig H Dafa'alla
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  2. George C Condon
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  3. Kirsty C Condon
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  4. Caroline E Phillips
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  5. Neil I Morrison
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  6. Li Jin
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  7. Matthew J Epton
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  8. Guoliang Fu
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  9. Luke Alphey
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Contributions

T.H.D. and G.C.C. contributed equally to this work. T.H.D., C.E.P. and L.J. designed and constructed the DNA constructs and assisted with the molecular analysis of transgenics. G.C.C. and K.C.C. created the transgenic Medflies and conducted most of the phenotypic and molecular analysis. N.I.M., M.J.E. and G.F. performed preliminary experiments and developed the marker system. L.A. conceived and supervised the project and wrote the paper with G.C.C. All authors discussed the results and commented on the manuscript.

Note: Supplementary information is available on the Nature Biotechnology website.

Corresponding author

Correspondence toLuke Alphey.

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Competing interests

Those authors affiliated with Oxitec Ltd. are employees or students of this company. Oxitec supplied salary and other support for the research program. Also, some of these authors have shares or share options in Oxitec Ltd. Both Oxitec Ltd. and Oxford University have one or more patents or patent applications related to the subject of this paper.

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Dafa'alla, T., Condon, G., Condon, K. et al. Transposon-free insertions for insect genetic engineering.Nat Biotechnol 24, 820–821 (2006). https://doi.org/10.1038/nbt1221

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