Overview

Also Known As:DEREG

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.

Donating Investigator

Tim Sparwasser, University Medical Center of the Johannes Gutenberg University Mainz

Genetic overview

Genetic Background	Generation

Tg(Foxp3-DTR/EGFP)23.2Spar

Allele Type
Transgenic (Inserted expressed sequence)

View Genetics

Research Applications

Research Tools
Immunology, Inflammation and Autoimmunity Research
Internal/Organ Research

View All Research Applications

Details

Detailed Description

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.

Development

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.

Expression Data

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.

Control Suggestions

Noncarrier
001026 BALB/cByJ

Additional Information

Considerations for Choosing Controls

Selected References

Jung S; Unutmaz D; Wong P; Sano G; De los Santos K; Sparwasser T; Wu S; Vuthoori S; Ko K; Zavala F; Pamer EG; Littman DR; Lang RA. 2002. In vivo depletion of CD11c(+) dendritic cells abrogates priming of CD8(+) T cells by exogenous cell-associated antigens. Immunity 17(2):211-20PubMed: 12196292 MGI: J:93488
Lahl K; Loddenkemper C; Drouin C; Freyer J; Arnason J; Eberl G; Hamann A; Wagner H; Huehn J; Sparwasser T. 2007. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J Exp Med 204(1):57-63PubMed: 17200412 MGI: J:125295

View All References

Genetics

Tg(Foxp3-DTR/EGFP)23.2Spar

Allele Symbol: Tg(Foxp3-DTR/EGFP)23.2Spar

Allele Name	transgene insertion 23.2, Tim Sparwasser
Allele Type	Transgenic (Inserted expressed sequence)
Allele Synonym(s)	DEREG; Tg(Foxp3-DTR/EGFP)1Ebe
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 expressionwithin the CD4+ T cell compartment, with the highest expression within the CD25+ subset.
Mutations Made By	Tim Sparwasser, Twincore Inst. for Infection Immunology

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Genetics Research
  - Tissue/Cell Markers: T cell specific surface marker
  - Tissue/Cell Markers
- Immunology, Inflammation and Autoimmunity Research
  - T cell deficiency
  - production of T cell lines and hybridomas
  - specific T cell deficiency
  - T cell specific surface marker
- Cancer Research
  - T cell deficiency
  - production of T cells and hybridoma
  - specific T cell deficiency
- Fluorescent Proteins
Immunology, Inflammation and Autoimmunity Research
- Immunodeficiency
  - specific T cell deficiency
  - T cell deficiency
- Autoimmunity
- Intracellular Signaling Molecules
- Inflammation
- CD Antigens, Antigen Receptors, and Histocompatibility Markers
  - genes regulating susceptibility to infectious disease and endotoxin
Internal/Organ Research
- Lymphoid Tissue Defects
  - T cell deficiency

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Tg(Foxp3-DTR/EGFP)23.2Spar/0
involves: C57BL/6NCrl

endocrine/exocrine gland phenotype

insulitis
- following diphtheria toxin treatment of neonates, mice exhibit insulitis and portal aggregates unlike similarly treated wild-type mice

growth/size/body region phenotype

enlarged spleen
- following diphtheria toxin treatment of neonates

integument phenotype

epidermal hyperplasia
- following diphtheria toxin treatment of neonates, the skin overlying the hyaline cartilage of the ear is thickened with epidermal hyperplasia and a dense lymphocyte infiltrate unlike in similarly treated wild-type mice

skin inflammation
- following diphtheria toxin treatment of neonates, the skin overlying the hyaline cartilage of the ear is thickened with epidermal hyperplasia and a dense lymphocyte infiltrate unlike in similarly treated wild-type mice

mammalian phenotype

immune system phenotype
- Normal - mice exhibit normal numbers and frequencies of CD25+CD4+ T regulatory cells

respiratory system phenotype

abnormal pulmonary alveolus morphology
- following diphtheria toxin treatment of neonates, alveoli are delicate and have thin walls compared to in similarly treated wild-type mice

lung inflammation
- following diphtheria toxin treatment of neonates, mice exhibit peribronchial and perivascular infiltrated unlike similarly treated wild-type mice

hematopoietic system phenotype

decreased regulatory T cell number
- following diphtheria toxin treatment, CD4+ T regulatory cells are decreased in adult and neonates compared to in similarly treated wild-type mice
- Normal - however, T regulatory cell numbers rebound after cessation of diphtheria treatment in adult mice but not neonates

enlarged spleen
- following diphtheria toxin treatment of neonates

increased spleen red pulp amount
- following diphtheria toxin treatment of neonates

increased spleen white pulp amount
- following diphtheria toxin treatment of neonates

immune system phenotype

decreased regulatory T cell number
- following diphtheria toxin treatment, CD4+ T regulatory cells are decreased in adult and neonates compared to in similarly treated wild-type mice
- Normal - however, T regulatory cell numbers rebound after cessation of diphtheria treatment in adult mice but not neonates

enlarged lymph nodes
- following diphtheria toxin treatment of neonates

enlarged spleen
- following diphtheria toxin treatment of neonates

increased inflammatory response
- diphtheria toxin treated-neonates exhibit massive inflammatory infiltrate in various organs that resembles the phenotype of Foxp3^sf hemizygotes

increased spleen red pulp amount
- following diphtheria toxin treatment of neonates

increased spleen white pulp amount
- following diphtheria toxin treatment of neonates

increased susceptibility to type IV hypersensitivity reaction
- following diphtheria toxin treatment, adult mice exhibit an enhanced and prolonged delayed-type hypersensitivity response compared with similarly treated wild-type mice

insulitis
- following diphtheria toxin treatment of neonates, mice exhibit insulitis and portal aggregates unlike similarly treated wild-type mice

lung inflammation
- following diphtheria toxin treatment of neonates, mice exhibit peribronchial and perivascular infiltrated unlike similarly treated wild-type mice

lymph node hyperplasia
- following diphtheria toxin treatment of neonates

skin inflammation
- following diphtheria toxin treatment of neonates, the skin overlying the hyaline cartilage of the ear is thickened with epidermal hyperplasia and a dense lymphocyte infiltrate unlike in similarly treated wild-type mice

References

Jung S; Unutmaz D; Wong P; Sano G; De los Santos K; Sparwasser T; Wu S; Vuthoori S; Ko K; Zavala F; Pamer EG; Littman DR; Lang RA. 2002. In vivo depletion of CD11c(+) dendritic cells abrogates priming of CD8(+) T cells by exogenous cell-associated antigens. Immunity 17(2):211-20PubMed: 12196292 MGI: J:93488
Lahl K; Loddenkemper C; Drouin C; Freyer J; Arnason J; Eberl G; Hamann A; Wagner H; Huehn J; Sparwasser T. 2007. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J Exp Med 204(1):57-63PubMed: 17200412 MGI: J:125295

Additional - Tg(Foxp3-DTR/EGFP)23.2Spar related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Tg(Foxp3-DTR/EGFP)23.2Spar
- Standard PCR:Tg(Foxp3-DTR/EGFP)23.2Spar
- Genotyping resources and troubleshooting
Breeding Considerations

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.
- Additional Breeding and Husbandry Support
Citation
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.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

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Licensing Information

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Also Known As:DEREG

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Tg(Foxp3-DTR/EGFP)23.2Spar

Allele Symbol: Tg(Foxp3-DTR/EGFP)23.2Spar

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Tg(Foxp3-DTR/EGFP)23.2Spar/0involves: C57BL/6NCrl

endocrine/exocrine gland phenotype

growth/size/body region phenotype

integument phenotype

mammalian phenotype

respiratory system phenotype

hematopoietic system phenotype

immune system phenotype

References

Additional - Tg(Foxp3-DTR/EGFP)23.2Spar related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Genotype: Tg(Foxp3-DTR/EGFP)23.2Spar/0
involves: C57BL/6NCrl