The Z/Edn2 knock-in gives constitutive expression of a nuclear-localized beta-galactosidase/neomycin resistance protein reporter (NLS-beta-geo) in the absence of Cre-mediated recombination. Following Cre-mediated recombination, the NLS-beta-geo coding region is excised and the downstream Edn2 coding region is expressed. This strain is useful in studies of retinal angiogenesis.
Jeremy Nathans, Johns Hopkins University
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) | Igs1 | intergenic site 1 |
Constitutive over-expression of endothelin-2 (Edn2) in the mouse retina perturbs vascular development by inhibiting endothelial cell migration across the retinal surface and subsequent endothelial cell invasion into the retina.
The Z/Edn2 knock-in, located 2 kb 5' of the ubiquitin b gene (the integration site of the Z/AP reporter), gives constitutive expression of a nuclear-localized beta-galactosidase/neomycin resistance protein reporter (NLS-beta-geo) in the absence of Cre-mediated recombination. Following Cre-mediated recombination, the NLS-beta-geo coding region is excised and the downstream Edn2 coding region is expressed.
When crossed with Sox2-Cre mice (see Stock No. 008454), Z/Edn;Sox2-Cre embryos recovered at E9.5-E10.5 are severely growth-retarded and display an open neural tube, an enlarged heart, and pooled blood in the head and thorax. Flatmount immunostaining for PECAM1 shows a nearly complete absence of vascular development. GFP is present in the targeting construct, but does not produce detectable fluorescence upon cre-mediated excision of the stop.
The following elements were inserted, in order, upstream of the Ubb locus by targeted recombination in (129X1/SvJ x 129S1/Sv)F1- Kitl+-derived R1 embryonic stem (ES) cells: CMV enhancer/beta-actin promoter, loxP site, beta-geo, 3 stop transcription sites, loxP site, Edn2 coding region, and IRES-GFP. This strain was maintained on a mixed C57BL/6 and 129 genetic background by the donating laboratory. GFP is present, but does not produce detectable fluorescence upon cre-mediated excision of the stop.
Expressed Gene | Bgeo, fusion of beta-galactosidase and neomycin phosphotransferase genes, E. coli |
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Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Edn2, endothelin 2, mouse, laboratory |
Site of Expression | GFP and Edn2 will be expressed in cells where promoters driving Cre recombinase are expressed |
Allele Name | targeted mutation 11, Jeremy Nathans |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) |
Allele Synonym(s) | Z/Edn2 |
Gene Symbol and Name | Igs1, intergenic site 1 |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | Bgeo, fusion of beta-galactosidase and neomycin phosphotransferase genes, E. coli |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Edn2, endothelin 2, mouse, laboratory |
Site of Expression | GFP and Edn2 will be expressed in cells where promoters driving Cre recombinase are expressed |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 11 |
Molecular Note | A site 5' of the Ubb gene was targeted with a construct composed of a CMV/beta-actin enhancer and promoter (CAG), a floxed beta-geo with three transcription termination sites and an Edn2 and IRES-GFP. Cre-mediated excision of the floxed beta-geo construct is required to achieve expression of the Edn2 and GFP cassette. |
Heterozygotes are viable and fertile. Homozygotes are unhealthy.
When using the Z/Edn2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #027109 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Igs1<tm11(CAG-Bgeo,-Edn2)Nat> |
Frozen Mouse Embryo | B6;129-Igs1<tm11(CAG-Bgeo -Edn2)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Igs1<tm11(CAG-Bgeo -Edn2)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Igs1<tm11(CAG-Bgeo -Edn2)Nat>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129-Igs1<tm11(CAG-Bgeo -Edn2)Nat>/J Frozen Embryo | $3373.50 |
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