These "depletion of regulatory T cell" (DEREG) BAC transgenic mice express a simian diphtheria toxin receptor-enhanced green fluorescent protein (DTR-eGFP) fusion protein under control of the endogenous forkhead box P3 promoter/enhancer regions on the BAC transgene. DTR-eGFP expression is observed in fully functional Foxp3+CD4+ regulatory T cell populations allowing fluorescent detection or diphtheria toxin-induced ablation Foxp3+ T reg cells.
Tim Sparwasser, University Medical Center of the Johannes Gutenberg University Mainz
Genetic Background | Generation |
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Allele Type |
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Transgenic (Inserted expressed sequence) |
Mice hemizygous for the "depletion of regulatory T cell" (DEREG) BAC transgene are viable and fertile, with expression of a simian diphtheria toxin receptor-enhanced green fluorescent protein (DTR-eGFP) fusion protein under control of the endogenous Foxp3 (forkhead box P3) promoter/enhancer regions on the BAC transgene. The donating investigator reports that transcription/translation from the BAC Foxp3 locus is disabled. These DEREG mice have DTR-eGFP expression in fully functional Foxp3+CD4+ regulatory T cell populations with no observed influence on regulation of the endogenous Foxp3 locus. Specifically, EGFP expression (via FACS) is observed mainly in CD25+CD4+ T cells, and allows specific detection of Foxp3+ regulatory T cell populations. The donating investigator also reports that EGFP expression is measurable by direct fluorescence. Diphtheria toxin (DT) administration results in ablation of Foxp3+CD4+ T reg cells with no apparent affect on CD25+ effector T cells. DT-induced depletion of Foxp3+CD4+ T reg cells is associated with enhanced and prolonged delayed-type hypersensitivity (DTH) responses and neonatal development of scurfy-like symptoms. The donating investigator reports that while homozygous females breed fine, homozygous males seem infertile.
In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.
The RP23-267C15 bacterial artificial chromosome (BAC) containing at least ten mouse genes was obtained. This BAC was modified by targeting a simian diphtheria toxin receptor-enhanced green fluorescent protein (DTR-eGFP) fusion protein and an SV40 polyA element into the first exon of the Foxp3 (forkhead box P3) locus: none of the other loci on the BAC were mutated. This modified ~150 kb BAC transgene was microinjected into the pronuclei of fertilized C57BL/6NCrl oocytes. Transgenic mice were identified and bred to C57BL/6J wildtype mice to establish the high transgene expressing founder line 23.2. These "depletion of regulatory T cell" (DEREG) mice were maintained on a C57BL/6J genetic background. Next, transgenic mice were backcrossed to BALB/cJ wildtype mice for 10 generations (five "speed-congenic" backcrosses and then five "conventional" backcrosses). After this, transgenic mice were backcrossed 2-3 generations to BALB/cByJ wildtype mice prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred to BALB/cByJ inbred mice for at least one generation to establish the colony.
Expressed Gene | DTR, Simian Diphtheria Toxin Receptor, |
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Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Selective DTR-eGFP expression is seen within the CD4+ T cell compartment, and highest expression within the CD25+ subset. |
Allele Name | transgene insertion 23.2, Tim Sparwasser |
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Allele Type | Transgenic (Inserted expressed sequence) |
Allele Synonym(s) | transgene insertion 23.2, Tim Sparwasser; Tg(Foxp3-DTR/EGFP)23.2Spar |
Gene Symbol and Name | Tg(Foxp3-DTR/EGFP)23.2Spar, transgene insertion 23.2, Tim Sparwasser |
Gene Synonym(s) | |
Promoter | Foxp3, forkhead box P3, mouse, laboratory |
Expressed Gene | DTR, Simian Diphtheria Toxin Receptor, |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Selective DTR-eGFP expression is seen within the CD4+ T cell compartment, and highest expression within the CD25+ subset. |
Strain of Origin | C57BL/6NCrl |
Chromosome | UN |
Molecular Note | The RP23-267C15 bacterial artificial chromosome (BAC) containing at least ten mouse genes was obtained. This BAC was modified by targeting a simian diphtheria toxin receptor-enhanced green fluorescent protein (DTR-eGFP) fusion protein and an SV40 polyA element into the first exon of the Foxp3 (forkhead box P3) locus: none of the other loci on the BAC were mutated. Two high expressing lines (16.1 and 23.2) were established. Analysis revealed selective DTR-eGFP expression within the CD4+ T cell compartment, with the highest expression within the CD25+ subset. |
Mutations Made By | Tim Sparwasser, Twincore Inst. for Infection Immunology |
When maintaining a live colony, hemizygous mice may be bred to wildtype (noncarrier) siblings or to BALB/cByJ inbred mice (Stock No. 001026). The donating investigator reports that while homozygous females breed fine, homozygous males seem infertile.
When using the DEREG mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #32049 in your Materials and Methods section.
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