B6.129S-Tcrd^{tm1.1(cre/ERT2)Zhu}/J

Stock number: 031679
Product name: TCRδ^CreER
Research areas: Cell Biology Research, Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

Tcrd^CreER knock-in mice express tamoxifen-inducible Cre recombinase under the direction of the T cell receptor delta chain (Tcrd) promoter in γδ T cells. These mice are useful for lineage tracking and genetic manipulation of the tissue resident γδ T cells.

Detailed description

The Tcrd^CreER knock-in allele has a tamoxifen-inducible Cre recombinase inserted downstream of the stop codon of the T cell receptor delta chain (Tcrd) gene. TCRD expression starts after T lineage commitment at the DN2 stage in the thymus and remains on in all mature γδ T cells. Mature αβ T cells do not contain the TCRD gene due to somatic deletion following TCRA gene recombination at the DP stage of T cell development in the thymus. Homozygous Tcrd^CreER mice are viable and fertile. When bred with mice containing a loxP-flanked sequence, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.

For example, when bred to Ai6 mice, which have Cre-inducible expression of an enhanced green fluorescent protein (ZsGreen1) (Stock No. 007906), tamoxifen-inducible Cre recombination results in expression of ZsGreen1 in the majority of γδ T cells in the skin, and a small fraction of CD4 or CD8 αβ T cells, but not in B cells, three days post tamoxifen treatment. Expression was also evident in γδ T cells in thymus, lymph nodes, Peyer’s patches, and liver.

Development

A targeting construct was designed to insert an internal ribosome entry site (IRES) and a cre/ERT2 fusion gene (Cre recombinase fused to a human estrogen receptor ligand binding domain) followed by a FRT-flanked neomycin resistance (neo) cassette, just downstream of the coding region of the T cell receptor delta chain (Tcrd) gene. This construct was electroporated into 129/Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to C57BL/6J mice. Offspring were bred to C57BL/6-Tg(CAG-flpe)2Arte mice to remove the neo cassette. Progeny were crossed to remove the flp-expressing transgene and the resulting Tcrd^CreER mice were bred to C57BL/6J inbred mice for at least 10 generations. Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Tcrd^{tm1.1(cre/ERT2)Zhu}
Name: targeted mutation 1.1, Yuan Zhuang
MGI accession ID: MGI:6117766
Generation method: Targeted
Attributes: Recombinase-expressing; Inducible
Synonyms: TCRd^CreER
Marker symbol: Tcrd
Marker name: T cell receptor delta chain
Marker synonyms: Tcr delta; Tcrdelta
Chromosome: 14
Strain of origin: 129S/SvEv
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: When bred with mice containing a loxP-flanked sequence, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.
Molecular note: A targeting construct was designed to insert an internal ribosome entry site (IRES) and a creERT2 fusion gene (Cre recombinase fused to a human estrogen receptor ligand-binding domain) followed by a frt-flanked neomycin resistance (neo) cassette, just downstream of the coding region of the T cell receptor delta chain (Tcrd) gene. This construct was electroporated into 129/sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to C57BL/6J mice. Offspring were bred to C57BL/6-Tg(CAG-Flpe)2Arte mice to remove the neo cassette. Progeny was crossed to remove the Flp-expressing transgene and the resulting mice were bred to C57BL/6J inbred mice for at least 10 generations.