B6.Cg-Wnt4^{tm3(EGFP/cre)Amc}/J

Stock number: 032490
Product name: Wnt4^GC, Wnt4GC, Wnt4-GC
Research areas: Developmental Biology Research, Research Tools

Description

These Wnt4^GC knock-in/knock-out mice express an eGFPCre, Enhanced Green Fluorescent Protein and Cre recombinase, fusion gene from the endogenous Wnt4 locus and may be useful in studies of nephrogenesis.

Detailed description

The extracellular signaling factor encoded by Wnt4 (wingless-type MMTV integration site family, member 4) plays an important role in the mesenchyme-to-epithelial transition during nephron development. The Wnt4^GC knock-in/knock-out allele both abolishes Wnt4 gene function and expresses an eGFPCre fusion protein (EGFP and Cre recombinase fusion protein) from the endogenous Wnt4 promoter/enhancer elements. Cre recombinase activity is detected in cap mesenchyme cells of the uretic bud (renal vesicle, comma head of the comma-shaped body, and median region of the S-shaped body). GFP expression is detected by immunolabeling in the mesonephros of 12.5 dpc embryos, in the gonad only of XX gonads, as well as in metanephric kidney of E15.5 embryos. Heterozygous mice are viable and fertile. Homozygous mice are not viable.

Development

A targeting vector was designed to insert an eGFPCre (GC) fusion protein coding sequence and an FRT site-flanked PGKneobpA selection cassette into the initiation codon of the Wnt4 (wingless-type MMTV integration site family, member 4) gene. The targeting vector contained an enhanced green fluorescent protein (EGFP) and a Cre recombinase fusion gene, an SV40 polyA signal, and an FRT site-flanked PGK-neo-bpA cassette. This construct was electroporated into (C57BL/6 x 129S4/SvJae)F1 derived v6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6J females. The mice were backcrossed to C57BL/6J for approximately 11 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Wnt4^{tm3(EGFP/cre)Amc}
Name: targeted mutation 3, Andrew P McMahon
MGI accession ID: MGI:4359657
Generation method: Targeted
Attributes: Recombinase-expressing
Synonyms: Wnt4^GC
Marker symbol: Wnt4
Marker name: wingless-type MMTV integration site family, member 4
Marker synonyms: SERKAL; WNT-4
Chromosome: 4
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Molecular note: The targeting vector was designed to insert an eGFP/cre (GC) fusion protein coding sequence and an Frt-flanked PGK-neo-polyA selection cassette into the initiation codon of the Wnt4 gene.