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Toxoplasma secreting Cre recombinase for analysis of host-parasite interactions

Abstract

We describe a Toxoplasma gondii strain that will permit the use of site-specific recombination to study the host-parasite interactions of this organism. This Toxoplasma strain efficiently injects a Cre fusion protein into host cells. In a Cre-reporter cell line, a single parasite invasion induced Cre-mediated recombination in 95% of infected host cells. By infecting Cre-reporter mice with these parasites, we also monitored host-cell infection in vivo.

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Figure 1: SeCreEt parasites.
Figure 2: SeCreEt parasites activate eGFP in reporter cell lines.
Figure 3: Analysis of SeCreEt parasites in vivo.

References

  1. Lewandoski, M. Nat. Rev. Genet. 2, 743–755 (2001).

    Article  CAS  Google Scholar 

  2. Cicin-Sain, L., Podlech, J., Messerle, M., Reddehase, M.J. & Koszinowski, U.H. J. Virol. 79, 9492–9502 (2005).

    Article  CAS  Google Scholar 

  3. Bradley, P.J. et al. J. Biol. Chem. 280, 34245–34258 (2005).

    Article  CAS  Google Scholar 

  4. Saeij, J.P. et al. Nature 445, 324–327 (2007).

    Article  CAS  Google Scholar 

  5. Neudeck, A. et al. Microbes Infect. 4, 581–590 (2002).

    Article  CAS  Google Scholar 

  6. Dubey, J.P., Sharma, S.P., Juranek, D.D., Sulzer, A.J. & Teutsch, S.M. Am. J. Vet. Res. 42, 1007–1010 (1981).

    CAS  PubMed  Google Scholar 

  7. Subauste, C.S. & Remington, J.S. Curr. Opin. Immunol. 5, 532–537 (1993).

    Article  CAS  Google Scholar 

  8. Gubbels, M.J., Striepen, B., Shastri, N., Turkoz, M. & Robey, E.A. Infect. Immun. 73, 703–711 (2005).

    Article  CAS  Google Scholar 

  9. Poupel, O., Boleti, H., Axisa, S., Couture-Tosi, E. & Tardieux, I. Mol. Biol. Cell 11, 355–368 (2000).

    Article  CAS  Google Scholar 

  10. Lodoen, M.B., Gerke, C. & Boothroyd, J.C. Cell Microbiol. 12, 55–66 (2010).

    Article  CAS  Google Scholar 

  11. Donald, R.G., Carter, D., Ullman, B. & Roos, D.S. J. Biol. Chem. 271, 14010–14019 (1996).

    Article  CAS  Google Scholar 

  12. Safran, M. et al. Mol. Imaging 2, 297–302 (2003).

    Article  CAS  Google Scholar 

  13. Saeij, J.P., Boyle, J.P., Grigg, M.E., Arrizabalaga, G. & Boothroyd, J.C. Infect. Immun. 73, 695–702 (2005).

    Article  CAS  Google Scholar 

  14. Vergunst, A.C. et al. Science 290, 979–982 (2000).

    Article  CAS  Google Scholar 

  15. Dranoff, G. et al. Proc. Natl. Acad. Sci. USA 90, 3539–3543 (1993).

    Article  CAS  Google Scholar 

  16. Pear, W., Scott, M. & Nolan, G.P. in Methods in Molecular Medicine: Gene Therapy Protocols (ed., Robbin, P.) 41–57 (Humana Press, Totowa, New Jersey, USA, 1997).

  17. Endo, T. & Yagita, K. J. Protozool. 37, 133–138 (1990).

    Article  CAS  Google Scholar 

  18. Liang, X. et al. Ann. Neurol. 64, 304–314 (2008).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank all members of the Boothroyd lab, especially M. Reese and S. Ravindran for technical help, J. Boyle for advice and G. Zeiner for suggesting the utility of a Toxoplasma strain that secretes Cre recombinase; G. Almogy for his help with the Cre-reporter cells; J.-F. Dubremetz (Université de Montpellier) for antibodies; M.-J. Gubbels (Boston College) for insightful comments and initial luciferase expression constructs; and N. Woodling (Stanford University) and members of the Andreasson lab for help with quantitative PCR. This research was supported by the US National Institutes of Health (NIH AI21423 and AI41014 to J.C.B.), the California HIV/AIDS Research Program of the University of California (F07-ST-201), the Walter and Idun Berry Fellowship and NIH (NS065116 to A.A.K.). M.B.L. was supported by the A.P. Giannini Foundation Medical Research Fellowship. H.M.B. and O.A. research was supported by NIH (AG009521, EB005011, AG020961 and AG024987), Muscular Dystrophy Association (grant 4320) and the Baxter Foundation.

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Authors

Contributions

A.E.F. and A.A.K. generated the SeCreEt parasites. A.A.K. analyzed the SeCreEt parasites. M.B.L. developed the plasmid for making toxofilin a partner for injecting fusion proteins into host cells. O.A. created the Cre-reporter cell line. H.M.B. oversaw the work of O.A., and J.C.B. oversaw the work of A.A.K., A.E.F. and M.B.L.

Corresponding author

Correspondence to John C Boothroyd.

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The authors declare no competing financial interests.

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Supplementary Figures 1–2 and Supplementary Table 1 (PDF 610 kb)

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Koshy, A., Fouts, A., Lodoen, M. et al. Toxoplasma secreting Cre recombinase for analysis of host-parasite interactions. Nat Methods 7, 307–309 (2010). https://doi.org/10.1038/nmeth.1438

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  • DOI: https://doi.org/10.1038/nmeth.1438

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