This Pax2Egfp knock-in/knockout strain has its Pax2 gene disrupted by an inserted EGFP sequence. These mice may be useful in applications related to the study of the developing intermediate mesoderm (kidney) and male/female urogenital epithelial structures, renal coloboma syndrome, and renal-specific stem cells.
Gregory R Dressler, University of Michigan
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, Null/Knockout) | Pax2 | paired box 2 |
The Pax2Egfp knock-in/knockout allele has an EGFP with polyA signal upstream of the endogenous paired box gene 2 (Pax2) translational start site; both abolishing endogenous gene function and placing EGFP expression under direction of the Pax2 promoter/enhancer regions. All mice homozygous for the Pax2Egfp allele exhibit exencephaly, complete renal agenesis, and die at birth. The donating investigator did not assess if any eye/ear abnormalities were present. Mice heterozygous for the Pax2Egfp allele are viable, fertile, and phenotypically indistinguishable from wildtype littermate controls (do not exhibit kidney structural defects). In both heterozygous and homozygous mice, EGFP expression (direct fluorescence) is observed in a spatial and temporal pattern consistent with the expected expression of endogenous Pax2. The EGFP expression of Pax2Egfp heterozygous mice is described in detail below.
At embryonic day (E)9.5, EGFP expression is observed in optic cup, otic vesicle, the midbrain-hindbrain junction, and developing nephric duct. By E10.5, EGFP positive cells of the intermediate mesoderm are segregated into epithelial duct cells and adjacent mesenchyme. By E11.5, EGFP is seen in the ureteric bud and condensing mesenchyme of the metanephric kidney, as well as Pax2-positive neurons that developed at this time along the entire neural tube. At later stages of development, kidney epithelia, ureters, and all the male and female specific epithelia derived from the intermediate mesoderm are EGFP positive. In newborn mice, EGFP expression is strong in the entire nephrogenic zone (nephron differentiation is still ongoing). In adult kidneys, EGFP expression is observed in the renal papilla. When comparing EGFP expression of mice heterozygous for the Pax2Egfp knockout allele (Stock No. 018438) to mice heterozygous for the Pax2Eneo hypomorphic allele (Stock No. 018437), there is no qualitative difference in the temporal and spatial pattern of EGFP, although the intensity of EGFP is higher in Pax2Egfp heterozygotes compared to Pax2Eneo heterozygotes.
The Pax2-Eneo targeting vector was designed to insert an enhanced green fluorescent protein sequence (EGFP; with polyA signal) and frt-flanked PGK-neo cassette upstream of the translational start site in exon 1 of the paired box gene 2 gene (Pax2). The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6 females to establish the Pax2Eneo colony. The donating investigator reports that to remove the frt-flanked PGK-neo cassette, some Pax2Eneo mice were bred to C57BL/6 animals expressing FLP recombinase. The resulting mice with EGFP and single frt site remaining upstream of the Pax2 coding region were identified. These Pax2Egfp mice were subsequently bred to wildtype mice for several generations (and the FLP gene was removed). Pax2Egfp mice on a mixed C57BL/6;129 genetic background were then sent to The Jackson Laboratory Repository. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish The Jackson Laboratory Repository colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | At embryonic day (E)9.5, EGFP expression is observed in optic cup, otic vesicle, the midbrain-hindbrain junction, and developing nephric duct. By E10.5, EGFP positive cells of the intermediate mesoderm are segregated into epithelial duct cells and adjacent mesenchyme. By E11.5, EGFP is seen in the ureteric bud and condensing mesenchyme of the metanephric kidney, as well as Pax2-positive neurons that developed at this time along the entire neural tube. At later stages of development, kidney epithelia, ureters, and all the male and female specific epithelia derived from the intermediate mesoderm are EGFP positive. In newborn mice, EGFP expression is strong in the entire nephrogenic zone (nephron differentiation is still ongoing). In adult kidneys, EGFP expression is observed in the renal papilla. |
Allele Name | targeted mutation 1.1, Gregory R Dressler |
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Allele Type | Targeted (Reporter, Null/Knockout) |
Allele Synonym(s) | Pax2Egfp |
Gene Symbol and Name | Pax2, paired box 2 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | At embryonic day (E)9.5, EGFP expression is observed in optic cup, otic vesicle, the midbrain-hindbrain junction, and developing nephric duct. By E10.5, EGFP positive cells of the intermediate mesoderm are segregated into epithelial duct cells and adjacent mesenchyme. By E11.5, EGFP is seen in the ureteric bud and condensing mesenchyme of the metanephric kidney, as well as Pax2-positive neurons that developed at this time along the entire neural tube. At later stages of development, kidney epithelia, ureters, and all the male and female specific epithelia derived from the intermediate mesoderm are EGFP positive. In newborn mice, EGFP expression is strong in the entire nephrogenic zone (nephron differentiation is still ongoing). In adult kidneys, EGFP expression is observed in the renal papilla. |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 19 |
Molecular Note | An EGFP and FRT-flanked neo cassette were inserted upstream of the coding sequence in exon 1. Flp-mediated recombination removed the neo cassette. |
All mice homozygous for the Pax2Egfp allele exhibit exencephaly, complete renal agenesis, and die at birth. When maintaining a live colony, heterozygous mice may be bred together or to wildtype mice from the colony.
When using the B6;129-Pax2tm1.1Gdr/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018438 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous or Wildtype for Pax2<tm1.1Gdr> |
Frozen Mouse Embryo | B6;129-Pax2<tm1.1Gdr>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Pax2<tm1.1Gdr>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Pax2<tm1.1Gdr>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129-Pax2<tm1.1Gdr>/J Frozen Embryo | $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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