The "interferon-gamma reporter with endogenous polyA transcript" (GREAT) allele has an IRES-eYFP reporter cassette inserted between the translational stop codon and 3' UTR/polyA tail of the interferon gamma (Ifng) gene. Thus, the bicistronic IFNγ-IRES-eYFP mRNA is under control of the endogenous IFNγ promoter/enhancer regions with proper regulation defined by the endogenous 3' UTR and polyA tail. These GREAT mice are a fluorescent reporter strain for studying innate immunity/adaptive immunity against viral and intracellular bacterial infections, for studying anti-tumor biology, as well for studying aberrant IFN-γ expression associated with autoinflammatory/autoimmune diseases.
Richard M. Locksley, Univ of California San Francisco-HHMI
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, No functional change) | Ifng | interferon gamma |
The "interferon-gamma reporter with endogenous polyA transcript" (GREAT) allele has an IRES-eYFP reporter cassette inserted between the translational stop codon and 3' UTR/polyA tail of the interferon gamma (Ifng) gene. Thus, the bicistronic IFNγ-IRES-eYFP mRNA is under control of the endogenous IFNγ promoter/enhancer regions with proper regulation defined by the endogenous 3' UTR and polyA tail. Expression of eYFP is observed by FACS/immunofluorescence microscopy in Ifng-expressing cell types; including innate immune responding cells (natural killer [NK] cells, natural killer T [NKT] cells) and cytotoxic T lymphocyte (CTL) effector cells of the adaptive immune response (CD4+ and CD8+T cells).
Of note, the donating investigator reports that these GREAT mice are a more useful tool than their IFNγ-IRES-YFP-BGHpolyA knockin (YETI) mice. Because the YETI allele has a bovine growth hormone polyA tail (rather than the endogenous IFN-γ polyA tail), homozygous YETI mice exhibit aberrant bicistronic IFNγ-IRES-YFP mRNA regulation that results in high IFN-γ blood levels and premature death. In contrast, homozygous GREAT mice are viable, fertile and normal in size, with proper transcript regulation defined by the endogenous IFN-γ 3' UTR/polyA tail.
A targeting vector was designed with a loxP-flanked neomycin resistance cassette, an internal ribosome entry site (IRES), and an enhanced yellow fluorescent protein (eYFP) sequence. This vector was inserted downstream of the endogenous translational stop codon and upstream of the 3' untranslated region (UTR) and polyA tail of the interferon gamma (Ifng) gene via electroporation into 129S4/SvJae-derived PrmCre embryonic stem (ES) cells. These PrmCre (or PC3) ES cells contain the Tg(Prm-cre)70Og transgene with the protamine promoter directing cre expression to the male germline. Correctly targeted ES cells were injected into recipient blastocysts. Chimeric males were bred to C57BL/6 females. The male offspring from this initial breeding lack the floxed neo cassette in their germline, and were bred to female C57BL/6 mice. Offspring with the IRES-eYFP cassette and without the Pmc-Cre transgene were identified to establish the colony of "interferon-gamma reporter with endogenous polyA transcript" (GREAT) mice. These GREAT mice were subsequently backcrossed to BALB/cJ mice for at least ten generations. Upon arrival at The Jackson Laboratory Repository, mice were bred to BALB/cJ inbred mice (Stock No. 000651) for at least one generation.
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
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Site of Expression | Cells activated to express Ifng will express EYFP. |
Allele Name | targeted mutation 3.1, Richard M Locksley |
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Allele Type | Targeted (Reporter, No functional change) |
Allele Synonym(s) | GREAT; Ifngtm3(EYFP)Lky |
Gene Symbol and Name | Ifng, interferon gamma |
Gene Synonym(s) | |
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
Site of Expression | Cells activated to express Ifng will express EYFP. |
Strain of Origin | 129S4/SvJae-Tg(Prm-cre)70Og |
Chromosome | 10 |
Molecular Note | A targeting vector was designed with a loxP-flanked neomycin resistance cassette, an internal ribosome entry site (IRES), and an enhanced yellow fluorescent protein (eYFP) sequence. This vector was inserted downstream of the endogenous translational stop codon and upstream of the 3' untranslated region (UTR). Cre-mediated recombination removed the neo cassette. |
Mutations Made By | Richard Locksley, Univ of California San Francisco-HHMI |
When maintaining a live colony, homozygous mice may be bred together.
When using the Great mouse strain in a publication, please cite the originating article(s) and include JAX stock #017580 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Ifng<tm3.1Lky> |
Frozen Mouse Embryo | C.129S4(B6)-Ifng<tm3.1Lky>/J | $2595.00 |
Frozen Mouse Embryo | C.129S4(B6)-Ifng<tm3.1Lky>/J | $2595.00 |
Frozen Mouse Embryo | C.129S4(B6)-Ifng<tm3.1Lky>/J | $3373.50 |
Frozen Mouse Embryo | C.129S4(B6)-Ifng<tm3.1Lky>/J | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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