B6.Cg-Tcra^tm1Mom Tg(TcrLCMV)327Sdz/TacMmjax

Stock number: 025408
Product name: TCR α-; P14 TCRVα2Vβ8
Research areas: Hematological Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

These mice carry a re-arranged T-cell receptor specific for LCMV peptide P14 and a Tcra KO that does not develop endogenous mature TCR alpha beta cells. This strain may be useful for studying CD8+ T cell activation and differentiation and as a source of donor cells for adoptive transfer for in vivo studies of inflammation, tolerance and anti-viral responses.

Detailed description

These mice carry a re-arranged T-cell receptor (Tcra-V2, Tcra-J TA31 / Tcrb-V8.1, Tcrb-D, Tcrb-J 2.4) specific for peptide P14 from the LCMV (lymphocytic choriomeningitis virus) in the context of H2-D^b. The LCMV P14 T Cell Receptor transgenic is combined with a Tcra KO that does not develop endogenous mature TCR alpha beta cells. Almost all the peripheral T cells are CD8+ and express the transgenic TCR. The mice are useful for in vitro studies of CD8+ T cell activation and differentiation to cytotoxic lymphocytes and as a source of donor cells for adoptive transfer into C57BL/6 mice for in vivo studies of inflammation, tolerance and anti-viral responses.

Development

Zinkernagel et al, established a set of LCMV-specific cytotoxic T-cell clones from lethally irradiated B10.BR/SgSn(H2^k reconstituted with T-cell depleted C57BL/10Sn (H2^b bone marrow cells. The rearranged TCR alpha (Tcra-V2 and Tcra-J TA31) and beta (Tcrb-V8.1, Tcrb-D, Tcrb-J2.4) chain gene sequences from clone P14 were incorporated into transgene constructs regulated by the H2^b promoter and co-injected into B6D2F2 fertilized embryos. 10-20 copies of both the alpha and beta transgenes co-integrated on the same chromosome in male founder number 327. Founder 327 was backcrossed to C57BL/6J for at least 10 generations prior to being intercrossed to make this a homozygous strain. Mice were transferred to Taconic in May of 1997 by Dr. B.J. Fowlkes and embryo transfer-derived into the NIAID Emerging Models colony.

The Tcra^tm1Mom allele was generated using a targeting vector containing a PGKneo cassette to disrupt exon 1, which contains the majority of the extracellular domain. The construct was electroporated into 129S2/SvPas-derived D3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6J mice for twelve generations.

In 1998, B6.Cg-Tg(TcrLCMV)327Sdz was crossed to B6.129S2-Tcra^tm1Mom. The line was then bred to obtain mice homozygous for both the Tcra null allele and the transgene followed by selection using test crosses and FACS analysis to identify mice exhibiting high levels of expression of the Vbeta 8.1, Va 2 transgene. In 2014 the line was transferred to the MMRRC at The Jackson Laboratory. Upon arrival, mice were bred to C57BL/6J for at least 1 generation to establish the colony.

Allele

Symbol: Tg(TcrLCMV)327Sdz
Name: transgene insertion 327, Birgit Ledermann
MGI accession ID: MGI:2665105
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(TcrLCMV)327Sdz
Marker name: transgene insertion 327, Birgit Ledermann
Chromosome: UN
Strain of origin: (C57BL/6 x DBA/2)F2
Expressed genes: Tcrb,T cell receptor beta chain,mouse, laboratory; Tcra,T cell receptor alpha chain,mouse, laboratory
Molecular note: The transgene carries both Tcra and Tcrb. Ten to twenty copies of the Tcr transgene were reported to have integrated on the same chromosome. At least two transgenic lines are known to exist. Line 327 is the representative line. The second line is 318.
General note: Transgenic mice carry a T-cell receptor (Tcra-V2, Tcra-J TA31 / Tcrb-V8.1, Tcrb-D, Tcrb-J 2.4) specific for the gp33 protein of LCMV (lymphocytic choriomeningitis virus), H2-D^b.

Allele

Symbol: Tcra^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857255
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Tcra
Marker name: T cell receptor alpha chain
Chromosome: 14
Strain of origin: 129S2/SvPas
Molecular note: A PGK-neo cassette was used to disrupt exon 1, which encodes the majority of the extracellular domain. The targeting event was confirmed by Southern analysis.