EGFP is expressed from the forkhead box P3 (Foxp3) locus in these mice. Foxp3 expression is not disrupted. Foxp3+ Treg cells can be identified in these mice by EGFP expression.
Bernard Malissen, Centre d'immunologie de Marseille Luminy
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Reporter) | Foxp3 | forkhead box P3 |
An IRES-EGFP cassette was placed in exon 11 in the 3' untranslated region of the X-linked forkhead box P3 (Foxp3) gene, without disrupting expression of the endogenous gene in this mutant mouse line. FOXP3 is a transcription factor necessary 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, EGFP expression is evident in FOXP3+ Treg cells. Homozygous Foxp3GFP mice are viable and fertile.
This new model from Dr Bernard Malissen of Centre d'Immunologie de Marseille-Luminy, France, targets the IRES-GFP cassette to exon 11, downstream of the endogenous Foxp3 STOP codon with no defects in FOXP3 or regulatory T cell function observed. This strain serves as an alternative to B6.Cg-Foxp3tm2Tch/J (Stock No. 006772) which targets the IRES-GFP cassette in intron 9 of the coding region.
The targeting vector was designed to insert (from 5' to 3') an internal ribosomal entry site (IRES) fused to an enhanced green fluorescent protein (EGFP) and a self-excising loxP-flanked ACN cassette into exon 11 of the X-linked forkhead box P3 (Foxp3) gene. The ACN cassette contained a neomycin resistance (Neo) gene and a testes-specific angiotensin-converting enzyme promoter-driven Cre recombinase gene (tACE-Cre). The construct was electroporated into C57BL/6-derived Bruce-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into FVB/N blastocysts and chimeric males were bred to C57BL/6J females. The donating investigator reported that the resulting Foxp3GFP mice were maintained on a C57BL/6J background (see SNP note below). Upon arrival mice at The Jackson Laboratory these mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
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/6N genetic background.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | Foxp3, forkhead box P3, mouse, laboratory |
Site of Expression | EGFP expression is observed in FOXP3+ regulatory T cells. |
Allele Name | targeted mutation 1, Bernard Malissen |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Foxp3.LuciDTR-4; FOXP3EGFP; Foxp3-GDL |
Gene Symbol and Name | Foxp3, forkhead box P3 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Foxp3, forkhead box P3, mouse, laboratory |
Site of Expression | EGFP expression is observed in FOXP3+ regulatory T cells. |
Strain of Origin | B6.Cg-Thy1a |
Chromosome | X |
Molecular Note | An IRES-EGFP cassette and a floxed cre-neo autodeleter casstte were inserted into exon 11, which lies in the downstream untranslated region. Passage through the male germ line-derived ES cells resulted in deletion of the cre-neo autodeleter cassette. The presence of the EGFP cassette does not interfer with expression of the gene locus. EGFP expression occurs in Foxp3+ regulatory T cells. |
Mutations Made By | Bernard Malissen, Centre d'immunologie de Marseille Luminy |
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 Foxp3EGFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #018628 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or Wild-type for Foxp3<tm1Mal> |
Frozen Mouse Embryo | B6.Cg-Foxp3<tm1Mal>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Foxp3<tm1Mal>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Foxp3<tm1Mal>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Foxp3<tm1Mal>/J Frozen Embryo | $3373.50 |
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