B6N.129S-Havcr2^tm1Bmed/J

Stock number: 021883
Product name: Tim3^mut
Research areas: Apoptosis Research, Cell Biology Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

The Tim3^mut mutant has a deletion of the terminal portion of the Tim3 cytoplasmic domain, resulting in defective CD8+ T cell effector function, enhanced negative regulatory activity, and absence of gal-9-mediated apoptosis.

Detailed description

The Tim3^mut targeted mutation h21883 as the terminal portion of the Tim3 cytoplasmic domain (amino acids 263-280) replaced with a zeomycin-resistance cassette, while preserving other regions of the cytoplasmic tail (including the highly conserved tyrosine kinase motif [mouse Y256]). The Tim3^mut mutation results in normal expression of RNA encoding the extracellular domain (normal surface expression of the Tim3 extracellular domain), but no expression of RNA encoding the deleted cytoplasmic domain region. Homozygous mice are viable and fertile, with normal size and appearance. Homozygous deletion of the Tim3 terminal cytoplasmic domain results in significant defect in CD8+ T cell effector function, but no signaling differences in CD4+ T cells. Tim3^mut homozygosity results in enhanced suppressive function/negative regulatory activity of Tim3, and absence of gal-9-mediated apoptosis. Tim3^mut homozygous mice infected with influenza A exhibit decreased IFN-γ RNA expression and decreased morbidity/mortality.

Development

The Tim3^mut targeting vector was designed to insert a zeomycin-resistance gene into exon 7 of the hepatitis A virus cellular receptor 2 gene (Havcr2 or Tim3). This mutation results in a deletion of the terminal portion of the cytoplasmic domain (amino acids 263-280), while preserving other regions of the cytoplasmic tail (including the highly conserved tyrosine kinase motif [mouse Y256]). The targeting construct was electroporated into 129S-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric males were bred with C57BL/6NCr females to establish the Tim3^mut colony. These Tim3^mut mice were subsequently bred to C57BL/6NCr mice for at least ten generations prior to sending to The Jackson Laboratory Repository in 2014. Upon arrival, sperm was cryopreserved. To generate the living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6NJ female mice (Stock No. 005304). The Tim3^mut mice were then bred together, to wildtype mice from the colony, or to C57BL/6NJ mice.

Allele

Symbol: Havcr2^tm1Bmed
Name: targeted mutation 1, Benjamin D Medoff
MGI accession ID: MGI:5447924
Generation method: Targeted
Attributes: Not Applicable
Marker symbol: Havcr2
Marker name: hepatitis A virus cellular receptor 2
Chromosome: 11
Strain of origin: 129S/SvEv
Molecular note: A neo cassette was inserted into exon 7 encoding the cytoplasmic domain (resulting in the deletion of amino acids 263 to 280). QPCR comfirmed reduced transcript expression of the portion encoding the cytoplasmic domain in splenocytes.