B6;129-Igs1^{tm11(CAG-Bgeo,-Edn2)Nat}/J

Stock number: 027109
Product name: Z/Edn2
Research areas: Research Tools

Description

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.

Detailed description

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.

Development

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.

Allele

Symbol: Igs1^{tm11(CAG-Bgeo,-Edn2)Nat}
Name: targeted mutation 11, Jeremy Nathans
MGI accession ID: MGI:5544083
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Igs1
Marker name: intergenic site 1
Chromosome: 11
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Expressed genes: Bgeo,fusion of beta-galactosidase and neomycin phosphotransferase genes,E. coli; Edn2,endothelin 2,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: GFP and Edn2 will be expressed in cells where promoters driving Cre recombinase are expressed
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.