The Tim3mut 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.
Benjamin D Medoff, Massachusetts General Hospital / Harvard Medical School
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Not Applicable) | Havcr2 | hepatitis A virus cellular receptor 2 |
The Tim3mut 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 Tim3mut 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. Tim3mut homozygosity results in enhanced suppressive function/negative regulatory activity of Tim3, and absence of gal-9-mediated apoptosis. Tim3mut homozygous mice infected with influenza A exhibit decreased IFN-γ RNA expression and decreased morbidity/mortality.
The Tim3mut 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 Tim3mut colony. These Tim3mut 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 Tim3mut mice were then bred together, to wildtype mice from the colony, or to C57BL/6NJ mice.
Allele Name | targeted mutation 1, Benjamin D Medoff |
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Allele Type | Targeted (Not Applicable) |
Allele Synonym(s) | Tim3mut |
Gene Symbol and Name | Havcr2, hepatitis A virus cellular receptor 2 |
Gene Synonym(s) | |
Strain of Origin | 129S/SvEv |
Chromosome | 11 |
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. |
When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony, or to C57BL/6NJ inbred mice (Stock No. 005304). Homozygous mice are also viable and fertile, and may be bred together.
When using the Tim3mut mouse strain in a publication, please cite the originating article(s) and include JAX stock #021883 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Havcr2<tm1Bmed> |
Frozen Mouse Embryo | B6N.129S-Havcr2<tm1Bmed>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N.129S-Havcr2<tm1Bmed>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N.129S-Havcr2<tm1Bmed>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6N.129S-Havcr2<tm1Bmed>/J Frozen Embryo | $3373.50 |
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