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


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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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.

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Correspondence to Thorsten Buch or Ari Waisman.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Disruption of myelin in the corpus callosum of DT-treated MOGi-cre/iDTR mice. (PDF 2884 kb)

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Figure 1: Scheme of the iDTR system.
Figure 2: Generation of ES cells with Cre-inducible DT sensitivity.
Figure 3: DT-mediated ablation of T cells in CD4-Cre/iDTR mice.
Figure 4: Improvement of ablation by longer and more frequent DT application.
Figure 5: DT-mediated ablation of oligodendrocytes in MOGi-Cre/iDTR mice.
Figure 6: DT-specific humoral immunity.