A targeting vector was designed to insert an internal ribosome entry site (IRES)-enhanced yellow fluorescent protein (eYFP) fusion protein, followed by a floxed neomycin (neo) resistance cassette, downstream of the endogenous translational stop codon of the interleukin 12b (Il12b) gene. These mice may be useful for visualizing active IL-12 in DCs and macrophages during immune response.
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) | Il12b | interleukin 12b |
A targeting vector was designed to insert an internal ribosome entry site (IRES)-enhanced yellow fluorescent protein (eYFP) fusion protein, followed by a floxed neomycin (neo) resistance cassette, downstream of the endogenous translational stop codon of the interleukin 12b (Il12b) gene. These homozygous YET40 mice are viable, fertile, and normal in size. IL-12b, or p40, binds with p35 to form the heterodimeric cytokine IL-12, after induction by toll like receptors (TLRs), in macrophages and dendritic cells (DC). YET40 mice show p40 and YFP expression in DC in draining lymph nodes, after inoculation with Listeria monocytogenes or lipopolysaccharide (LPS), and promote Th1 differentiation. These mice may be useful for visualizing active IL-12 in DCs and macrophages during immune response.
A targeting vector was designed to insert an internal ribosome entry site (IRES)-enhanced yellow fluorescent protein (eYFP) fusion protein, followed by a floxed neomycin (neo) resistance cassette, downstream of the endogenous translational stop codon of the interleukin 12b (Il12b) gene. The construct was electroporated into 129S4/SvJae-derived PC3 embryonic stem (ES) cells. These PC3 cells contain a transgene expressing Cre recombinase under control of the protamine promoter (Tg(Prm-cre)700g), to delete the neo cassette in the male germline. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females. The male offspring from this initial breeding lack the floxed neo cassette and were bred to female C57BL/6 mice to establish a colony of YET40 mice. These mutant mice were backcrossed to BALB/cJ mice for at least 10 generations. Upon arrival at The Jackson Laboratory, mice were bred to BALB/cJ (Stock No. 000664) for at least one generation.
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
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Site of Expression | EYFP is expressed in dendritic cell (DC) and macrophage lineage cells that produce Il12b (p40) following stimulation with toll-like receptor (TLR) ligands both in vivo and in vitro. |
Allele Name | targeted mutation 1.1, Richard M Locksley |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | IL-12/p40 y; Il12btm1(EYFP)Lky; p40 (y); p40-IRES-eYFP; yet40 |
Gene Symbol and Name | Il12b, interleukin 12b |
Gene Synonym(s) | |
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
Site of Expression | EYFP is expressed in dendritic cell (DC) and macrophage lineage cells that produce Il12b (p40) following stimulation with toll-like receptor (TLR) ligands both in vivo and in vitro. |
Strain of Origin | 129S4/SvJae-Tg(Prm-cre)70Og |
Chromosome | 11 |
Molecular Note | A targeting vector was designed to insert an internal ribosomal entry site (IRES)-enhanced yellow fluorescent protein (EYFP; Clontech) sequence (followed by a bovine growth hormone poly(A) signal and a loxP site flanked neomycin resistance gene cassette) downstream of the stop codon in exon 8 of the endogenous gene. The construct was electroporated into ES cells that contain the Tg(Prm-cre)70Og transgene that effects the removal of the neo cassette through cre-mediated recombination in the male germline. |
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 p40-IRES-eYFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #015864 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Il12b<tm1Lky> |
Frozen Mouse Embryo | C.129S4(B6)-Il12b<tm1.1Lky>/J | $2595.00 |
Frozen Mouse Embryo | C.129S4(B6)-Il12b<tm1.1Lky>/J | $2595.00 |
Frozen Mouse Embryo | C.129S4(B6)-Il12b<tm1.1Lky>/J | $3373.50 |
Frozen Mouse Embryo | C.129S4(B6)-Il12b<tm1.1Lky>/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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