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Site-specific transgenesis by Cre-mediated recombination in Drosophila

Abstract

Transposons such as P elements are routinely used to stably transfer exogenous DNA (transgenes) into the Drosophila genome. Transgene insertion events, however, are essentially random and are subject to 'position effects' from nearby endogenous regulatory elements. Here we describe a microinjection-based system that uses Cre-mediated recombination to insert transgenes into precise genomic 'landing sites'. The system is simple and efficient, and will permit precise comparisons between multiple transgenic constructs.

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Figure 1: A strategy for RMCE in Drosophila.
Figure 2: Cre-mediated RMCE.
Figure 3: Precise comparisons of two reporter genes.

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References

  1. Barolo, S., Castro, B. & Posakony, J.W. Biotechniques 36, 436–440, 442 (2004).

    Article  CAS  Google Scholar 

  2. Cai, H.N., Zhang, Z., Adams, J.R. & Shen, P. Development 128, 4339–4347 (2001).

    CAS  PubMed  Google Scholar 

  3. Zhou, J. & Levine, M. Cell 99, 567–575 (1999).

    Article  CAS  Google Scholar 

  4. Siegal, M.L. & Hartl, D.L. Genetics 144, 715–726 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Siegal, M.L. & Hartl, D.L. Proc. Natl. Acad. Sci. USA 95, 15513–15518 (1998).

    Article  CAS  Google Scholar 

  6. Baer, A. & Bode, J. Curr. Opin. Biotechnol. 12, 473–480 (2001).

    Article  CAS  Google Scholar 

  7. Golic, M.M., Rong, Y.S., Petersen, R.B., Lindquist, S.L. & Golic, K.G. Nucleic Acids Res. 25, 3665–3671 (1997).

    Article  CAS  Google Scholar 

  8. Groth, A.C., Fish, M., Nusse, R. & Calos, M.P. Genetics 166, 1775–1782 (2004).

    Article  CAS  Google Scholar 

  9. Chen, Z.Y., He, C.Y., Ehrhardt, A. & Kay, M.A. Mol. Ther. 8, 495–500 (2003).

    Article  CAS  Google Scholar 

  10. Hoess, R.H., Wierzbicki, A. & Abremski, K. Nucleic Acids Res. 14, 2287–2300 (1986).

    Article  CAS  Google Scholar 

  11. Lee, G. & Saito, I. Gene 216, 55–65 (1998).

    Article  CAS  Google Scholar 

  12. Langer, S.J., Ghafoori, A.P., Byrd, M. & Leinwand, L. Nucleic Acids Res. 30, 3067–3077 (2002).

    Article  CAS  Google Scholar 

  13. Albert, H., Dale, E.C., Lee, E. & Ow, D.W. Plant J. 7, 649–659 (1995).

    Article  CAS  Google Scholar 

  14. Small, S. Methods Enzymol. 326, 146–159 (2000).

    Article  CAS  Google Scholar 

  15. Small, S., Blair, A. & Levine, M. EMBO J. 11, 4047–4057 (1992).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank M. Siegal for the pMLS104 Cre helper plasmid used in these experiments, V. Vasisht for plasmids containing loxP sequences, D. Papatsenko for loxP-y-loxP transgenic flies, M. Wernet for advice on the inverted PCR protocol, and M. Siegal and P. Struffi for comments on the manuscript. L.K. was supported by the 2004 Howard Hughes Medical Institute summer program for high school teachers. This work was supported by grant RO1 GM 51946 from the National Institutes of Health, and conducted in a facility constructed with support from Research Facilities Improvement Grant C06 RR-15518-01 from the National Center for Research Resources, National Institutes of Health.

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Correspondence to Stephen Small.

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Supplementary information

Supplementary Table 1

Cre-mediated RMCE in transgenic Drosophila. (PDF 94 kb)

Supplementary Table 2

Transgene excision tests. (PDF 96 kb)

Supplementary Methods (PDF 131 kb)

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Oberstein, A., Pare, A., Kaplan, L. et al. Site-specific transgenesis by Cre-mediated recombination in Drosophila. Nat Methods 2, 583–585 (2005). https://doi.org/10.1038/nmeth775

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