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A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration

Nature Methods volume 2, pages 419426 (2005) | Download Citation

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Abstract

A new system for lineage ablation is based on transgenic expression of a diphtheria toxin receptor (DTR) in mouse cells and application of diphtheria toxin (DT). To streamline this approach, we generated Cre-inducible DTR transgenic mice (iDTR) in which Cre-mediated excision of a STOP cassette renders cells sensitive to DT. We tested the iDTR strain by crossing to the T cell– and B cell–specific CD4-Cre and CD19-Cre strains, respectively, and observed efficient ablation of T and B cells after exposure to DT. In MOGi-Cre/iDTR double transgenic mice expressing Cre recombinase in oligodendrocytes, we observed myelin loss after intraperitoneal DT injections. Thus, DT crosses the blood-brain barrier and promotes cell ablation in the central nervous system. Notably, we show that the developing DT-specific antibody response is weak and not neutralizing, and thus does not impede the efficacy of DT. Our results validate the use of iDTR mice as a tool for cell ablation in vivo.

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Acknowledgements

We thank S. Becker and C. Uthoff-Hachenberg for technical assistance. We also thank J. van de Water, I. Parvanova and A. Croxford for critically reading the manuscript. This work was funded by Deutsche Forschungsgemeinschaft grants WA 1601/1-1 to A.W. and BU 1410/1-1 to T.B. T.B. was a fellow of the International Graduate School of Functional Genomics and Genetics at the Institute for Genetics, Cologne. F.L.H. was supported by the Stammbach foundation. S.J. is a Scholar of the Benoziyo Center for Molecular Medicine.

Author information

Author notes

    • Ari Waisman

    Present address: I. Medical Department, Johannes Gutenberg-University Mainz, 55131 Mainz, Germany.

Affiliations

  1. Laboratory for Molecular Immunology, Institute for Genetics, University of Cologne, D-50931 Cologne, Germany.

    • Thorsten Buch
    • , Christine Tertilt
    • , Tobias J A J Heinen
    • , F Thomas Wunderlich
    •  & Ari Waisman
  2. Institute of Neuropathology, University Hospital Zurich, CH-8091 Zurich, Switzerland.

    • Frank L Heppner
  3. Research Department, Kantonal Hospital St. Gallen, 9007 St. Gallen, Switzerland.

    • Marcel Kremer
  4. Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.

    • Steffen Jung

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The authors declare no competing financial interests.

Corresponding authors

Correspondence to Thorsten Buch or Ari Waisman.

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    Supplementary Fig. 1

    Disruption of myelin in the corpus callosum of DT-treated MOGi-cre/iDTR mice.

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https://doi.org/10.1038/nmeth762

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