Strain Detail Sheet

Strain Name    :

C.B6-Tg(Foxp3-DTR/EGFP)23.2Spar/Mmjax

Stock Number :

032049-JAX

Gene Information

Gene Details [Including genotyping protocols]

(provided by MGI)
Transgene: Tg(Foxp3-DTR/EGFP)23.2Spar
Name: transgene insertion 23.2, Tim Sparwasser
Alteration at locus: Transgenic
Reporter: EGFP (Jelly Fish)
Name: Enhanced Green Fluorescent Protein
Alteration at locus: Transgenic
Promoter: Foxp3
Name: forkhead box P3
Alteration at locus: Transgenic
Transgene: DTR (Simian)
Name: diphtheria toxin receptor
Alteration at locus: Transgenic

Genetic Alterations:
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.

Genotype Determination:

ES Cell Line: Not Applicable

Strain Description [Including phenotype, strain background, strain development and suggested control mice]

Phenotype

Homozygous: The donating investigator reports that while homozygous females breed fine, homozygous males seem infertile.

Hemizygous: 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.


Mammalian Phenotype Terms:(provided by MGI)      Extend all MPTs
      assigned by genotype

The following phenotype information may relate to one or more alleles on a genetic background differing from this MMRRC strain.
Tg(Foxp3-DTR/EGFP)23.2Spar/-
        involves: C57BL/6NCrl
  • immune system phenotype
    • immune system phenotype (MGI Ref ID J:125295)
      • mice exhibit normal numbers and frequencies of CD25+CD4+ T regulatory cells
      • decreased regulatory T cell number (MGI Ref ID J:125295)
        • following diphtheria toxin treatment, CD4+ T regulatory cells are decreased in adult and neonates compared to in similarly treated wild-type mice
        • however, T regulatory cell numbers rebound after cessation of diphtheria treatment in adult mice but not neonates
      • enlarged lymph nodes (MGI Ref ID J:125295)
        • following diphtheria toxin treatment of neonates
        • lymph node hyperplasia (MGI Ref ID J:125295)
          • following diphtheria toxin treatment of neonates
      • enlarged spleen (MGI Ref ID J:125295)
        • following diphtheria toxin treatment of neonates
      • increased inflammatory response (MGI Ref ID J:125295)
        • diphtheria toxin treated-neonates exhibit massive inflammatory infiltrate in various organs that resembles the phenotype of Foxp3sf hemizygotes
        • insulitis (MGI Ref ID J:125295)
          • following diphtheria toxin treatment of neonates, mice exhibit insulitis and portal aggregates unlike similarly treated wild-type mice
        • lung inflammation (MGI Ref ID J:125295)
          • following diphtheria toxin treatment of neonates, mice exhibit peribronchial and perivascular infiltrated unlike similarly treated wild-type mice
        • skin inflammation (MGI Ref ID J:125295)
          • 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
      • increased spleen red pulp amount (MGI Ref ID J:125295)
        • following diphtheria toxin treatment of neonates
      • increased spleen white pulp amount (MGI Ref ID J:125295)
        • following diphtheria toxin treatment of neonates
      • increased susceptibility to type IV hypersensitivity reaction (MGI Ref ID J:125295)
        • following diphtheria toxin treatment, adult mice exhibit an enhanced and prolonged delayed-type hypersensitivity response compared with similarly treated wild-type mice
  • endocrine/exocrine gland phenotype
    • insulitis (MGI Ref ID J:125295)
      • following diphtheria toxin treatment of neonates, mice exhibit insulitis and portal aggregates unlike similarly treated wild-type mice
  • respiratory system phenotype
    • abnormal pulmonary alveolus morphology (MGI Ref ID J:125295)
      • following diphtheria toxin treatment of neonates, alveoli are delicate and have thin walls compared to in similarly treated wild-type mice
    • lung inflammation (MGI Ref ID J:125295)
      • 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 (MGI Ref ID J:125295)
      • following diphtheria toxin treatment, CD4+ T regulatory cells are decreased in adult and neonates compared to in similarly treated wild-type mice
      • however, T regulatory cell numbers rebound after cessation of diphtheria treatment in adult mice but not neonates
    • enlarged spleen (MGI Ref ID J:125295)
      • following diphtheria toxin treatment of neonates
    • increased spleen red pulp amount (MGI Ref ID J:125295)
      • following diphtheria toxin treatment of neonates
    • increased spleen white pulp amount (MGI Ref ID J:125295)
      • following diphtheria toxin treatment of neonates
  • integument phenotype
    • epidermal hyperplasia (MGI Ref ID J:125295)
      • 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 (MGI Ref ID J:125295)
      • 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

Strain of Origin: C57BL/6NCrl

Strain genetic background: C.B6

Strain Development: The modified BAC transgene was microinjected into the pronuclei of fertilized C57BL/6NCrl oocytes. Transgenic mice were identified and bred to C57BL/6J wild-type 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 wild-type 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 wild-type mice prior to arrival at MMRRC. Upon arrival, mice were bred to BALB/cByJ inbred mice for at least one generation to establish the colony.

The allele in this mutant line is on a genetic background different from that on which the allele was first characterized.

Suggested Control Mice: Wild-type littermates

Research Applications

    Strain Origin

    Donor: Tim Sparwasser, Ph.D., Twincore, Zentrum fur Esperimentelle und Klin. Infektionsforschund GmbH

    Primary Reference:

    Lahl K; Loddenkemper C; Drouin C; Freyer J; Arnason J; Eberl G; Hamann A; Wagner H; Huehn J; Sparwasser T, Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease., J Exp Med 2007 Jan 22;204(1):57-63 (Medline PMID: 17200412)

    Colony and Husbandry Information

    Special Considerations

    When maintaining a live colony, hemizygous mice may be bred to wildtype (noncarrier) siblings or to BALB/cByJ inbred mice. The donating investigator reports that while homozygous females breed fine, homozygous males seem infertile.

    Health Status Report

    Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email csmmrrc@jax.org.

    Order Request Information

    Availability Level

    Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

    Conditions of Distribution [Including applicable technology transfer agreements]

    Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

    The donor or their institution limits the distribution to non-profit institutions only.

    Fees

    Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

    Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
    MMRRC Item # - Description Distribution
    Fee/unit (US $)
    Units Notes
    032049-JAX-RESUSLitter recovered from cryo-archive
    $2,022.00
    Non-Profit
    Litter Recovered litter1; additional fees for any special requests.
    032049-JAX-SPERMCryo-preserved spermatozoa
    $437.00
    Non-Profit
    Aliquot Approximate quantity.2

    1 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

    2 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.

    3 An aliquot contains a sufficient number of embryos (in one or more vials and based on the transfer success rate of the MMRRC facility) to transfer to at least two recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

    To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section above). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.



    To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.



    The MMRRC is a collaborative effort, funded by grants from DPCPSI of the NIH.

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