C57BL/6N-Tg(CD2-Tcra,-Tcrb)1Ybe/Mmjax

Stock number: 040048
Product name: SA1-Tg
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

SA1-Tg mice express T cell receptor α and β chains, driven by a human CD2 promoter, that recognize components of the common human skin commensal, Staphylococcus aureus.

Note that the donating investigator has also made an SA2-Tg line available as MMRRC RRID:072060.

Detailed description

SA1-transgenic (Tg) mice express T cell receptor α and β chains, driven by a human CD2 promoter, that recognize components of the common human skin commensal, Staphylococcus aureus. Hemizygous mice are viable and fertile with no overt phenotype. The donating laboratory has not attempted to generate homozygous mice. The SA1-Tg and SA2-Tg mouse lines (MMRRC RRID:072060) are S. aureus-specific transgenic mice where all CD4⁺ T cells are specific for components of S. aureus 42F02. However, each strain has a unique TCR construct and likely will not recognize the same S. aureus antigen. These mice may be useful for tracking S. aureus-specific CD4⁺ T cells in vivo.

Development

To generate the SA1-transgene (Tg), Foxp3-GFP reporter mice on a C57BL/6 background, were topically associated with S. aureus (42F02), and 4 weeks after the first association, mice were infected with S. aureus (42F02) by an intradermal injection in the ear to recall the S. aureus-specific T cells. An s. aureus-specific T cell suspension was obtained from the ear skin-draining lymph nodes and co-cultured with S. aureus loaded dendritic cells for several weeks. FoxP3^-D4⁺CCR6⁺ T cells were FACSorted from the in vitro culture and subjected to single-cell sequencing of TCRα and TCRβ chains. Clonal TCR pairs (differing from the TCR pair used for creating SA2-Tg mice, MMRRC RRID:072060) were identified and used in a hybridoma reconstitution screening assay to identify S. aureus-reactive TCR heterodimers. A single S. aureus-specific TCR pair was cloned into a human CD2-expression vector. The resulting SA1-Tg construct was injected into the pronuclei of fertilized C57BL/6Ncr oocytes. Founder line 1 was established and backcrossed at least 6 generations to C57BL/6NTac by the donating laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes.

Of note, the transgene insertion site and/or copy number are not known.

Allele

Symbol: Tg(CD2-Tcra,-Tcrb)1Ybe
Name: transgene insertion 1, Yasmine Belkaid
MGI accession ID: MGI:7782515
Generation method: Transgenic
Attributes: Inserted expressed sequence
Synonyms: SA1-Tg; Tg(CD2-Tcr*)1Ybe
Marker symbol: Tg(CD2-Tcra,-Tcrb)1Ybe
Marker name: transgene insertion 1, Yasmine Belkaid
Chromosome: UN
Strain of origin: C57BL/6NCr
Molecular note: Clonal TCR pairs were identified and used in a hybridoma reconstitution screening assay to identify S. aureus-reactive TCR heterodimers from mice infected with S. aureus (42F02) by an intradermal injection in the ear. A single S. aureus-specific TCR pair was cloned into a human CD2-expression vector. Founder line 1 was established.