C57BL/6-Trdc^tm1Mal/J

Stock number: 016941
Product name: Tcrd-H2BEGFP
Research areas: Research Tools

Description

EGFP expression in these targeted reporter mice is seen in CD3ε⁺ TCRγδ⁺ intraepithelial lymphocytes in the small intestines, and TCRγδ cells in the spleen and lymph nodes. These mice may be useful for visualizing and studying the development of adult thymic γδ T cells and the biology of peripheral γδ T cells.

Detailed description

Tcrd-H2BEGFP mutant mice contain an internal ribosome entry site (IRES), and a human histone H2B fused to an enhanced green fluorescent protein (EGFP) downstream of the internal stop codon of the T-cell receptor delta, constant region (Trdc) gene. Homozygotes are viable, fertile, and normal in size. EGFP expression is evident in all CD3ε⁺ TCRγδ⁺ intraepithelial lymphocytes (IELs) in the small intestines, and TCRγδ cells in the spleen and lymph nodes. These mice may be useful for visualizing and studying the development of adult thymic γδ T cells and the biology of γδ T cells in the periphery.

The donating investigator reports that for visualizing γδ T cells in secondary lymphoid organs or other tissues containing a large excess of αβ T cells over γδ T cells, the use of F1 heterozygotes from Tcrd-H2BEGFP mice and B6.129S2-Tcra^tm1Mom/J (Stock No. 002116) will genetically exclude highly fluorescent false-positive cells.

Development

The Tcrd-H2BEGFP targeting vector was designed to contain an internal ribosome entry site (IRES), and a human histone H2B fused to an enhanced green fluorescent protein (EGFP), followed by a self-excising loxP-flanked ACN cassette (itself containing a neomycin resistance (Neo) gene and a testes-specific angiotensin-converting enzyme promoter-driven Cre recombinase gene), which self-excises during transmission in the male chimera. This construct was inserted downstream of the internal stop codon of the T-cell receptor delta, constant region (Trdc) gene. This construct was injected into C57BL/6-derived Bruce-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into FVB blastocysts, and the resulting chimeric males were bred to C57BL/6J females (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) 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, 1 of 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 mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Trdc^tm1Mal
Name: targeted mutation 1, Bernard Malissen
MGI accession ID: MGI:3665255
Generation method: Targeted
Attributes: Reporter; Inserted expressed sequence; Humanized sequence
Marker symbol: Trdc
Marker name: T cell receptor delta, constant region
Chromosome: 14
Strain of origin: B6.Cg-Thy1^a
Expressed genes: GFP,Green Fluorescent Protein,; H2B-eGFP,Histone H2B-enhanced Green Fluorescent Protein,
Site of expression: EGFP is expressed in intraepithelial T-cells in the small intestine, spleen and lymph nodes.
Molecular note: An expression cassette encoding an internal ribosomal entry site (IRES) and a fusion of human histone H2B and enhanced green fluorescence protein (EGFP) was inserted in the 3' untranslated region.