Foxp3DTR mutant mice express knocked-in human diphtheria toxin receptor and EGFP genes from the Foxp3 locus--without disrupting expression of the endogenous Foxp3 gene. These mice may be useful for regulatory T cell visualizing and ablation.
Alexander Rudensky, Memorial Sloan Kettering Cancer Center
Genetic Background | Generation |
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N8+pN2F1
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, Inserted expressed sequence) | Foxp3 | forkhead box P3 |
Foxp3DTR mutant mice contain an internal ribosome entry site (IRES), a human diphtheria toxin receptor (DTR), and an enhanced green fluorescent protein (EGFP) downstream of the internal stop codon of the forkhead box P3 (Foxp3) gene. FOXP3 is a transcription factor required for the development and function of regulatory T (T reg) cells, which are required for suppression of self-reactive T cells and prevention of some autoimmune diseases. In these mice, DTR-EGFP expression is evident in FOXP3+ Treg cells. Diphtheria toxin (DT) administration results in ablation of Treg cells in thymus, lymph nodes, and spleen 2 days after injection, with cell numbers rebounding 10-15 days post-injection. Neonates injected with DT daily all die from lymphoproliferative disease, indicated by failure to thrive, lack of mobility, ventral skin lesions, hunched posture, and conjunctivitis, within 27 days of birth. With the same DT treatment, adults exhibit a more rapid development of autoimmune disease and die within 3 weeks. These mice show an increase in CD4+ T cell activation, as well as an increase in the number of B cells, macrophages, granulocytes, natural killer, and dendritic cells in the spleen and lymph nodes. Homozygous females and hemizygous males are viable, fertile, and normal in size. These mice may be useful for visualizing and specifically eliminating regulatory T cells, and for studying autoimmunity and immune dysfunction.
The Foxp3DTR targeting vector was designed with an internal ribosome entry site (IRES), a human diphtheria toxin receptor (DTR), and an enhanced green fluorescent protein (EGFP), followed by a frt-flanked neomycin (neo) resistance cassette inserted downstream of the internal stop codon of the X-linked forkhead box P3 (Foxp3) gene. This construct was injected into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The donating investigator reported that the resulting mice were backcrossed to C57BL/6NTac mice for at least 8 generations (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6 genetic background.
Expressed Gene | DTR/EGFP, Simian Diphtheria Toxin Receptor; Enhanced Green Fluorescent Protein, |
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Site of Expression | DTR-EGFP expression is evident in regulatory T cells. |
Allele Name | targeted mutation 3, Alexander Y Rudensky |
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Allele Type | Targeted (Reporter, Inserted expressed sequence) |
Allele Synonym(s) | targeted mutation 3, Alexander Y Rudensky; Foxp3tm3(DTR/GFP)Ayr |
Gene Symbol and Name | Foxp3, forkhead box P3 |
Gene Synonym(s) | AIID; sf; DIETER; IPEX; PIDX; scurfin; XPID; JM2; scurfy; RGD1562112 |
Expressed Gene | DTR/EGFP, Simian Diphtheria Toxin Receptor; Enhanced Green Fluorescent Protein, |
Site of Expression | DTR-EGFP expression is evident in regulatory T cells. |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | X |
Molecular Note | A targeting vector was designed to insert an IRES-DTR-GFP with a floxed neo downstream of exon 11. |
Mutations Made By | Alexander Rudensky, Memorial Sloan Kettering Cancer Center |
This mutant allele is located on the X chromosome. When maintaining a live colony, homozygous females may be bred with hemizygous males.
When using the Foxp3DTR mouse strain in a publication, please cite the originating article(s) and include JAX stock #016958 in your Materials and Methods section.
Service | Genotype | Price |
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X linked - Heterozygous females and wildtype males for Foxp3<tm3Ayr> |
We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.
Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.
Frozen Mouse Embryo | B6.129(Cg)-Foxp3<tm3(DTR/GFP)Ayr>/J | $2595.00 |
Frozen Mouse Embryo | B6.129(Cg)-Foxp3<tm3(DTR/GFP)Ayr>/J | $2595.00 |
Frozen Mouse Embryo | B6.129(Cg)-Foxp3<tm3(DTR/GFP)Ayr>/J | $3373.50 |
Frozen Mouse Embryo | B6.129(Cg)-Foxp3<tm3(DTR/GFP)Ayr>/J | $3373.50 |
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