B6;129S-Wnt5a^tm1.1Krvl/J

Stock number: 026626
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

Wnt5a^fl/fl floxed mice may be useful for studying the branching and extension of sympathetic axons in the peripheral nervous system.

Detailed description

Wnt5a^fl/fl floxed mice possess loxP sites flanking exon 2 of the wingless-type MMTV integration site family, member 5A (Wnt5a) gene. WNT5A is a member of the Wnt family of secreted proteins expressed in sympathetic neurons during target innervation. It is a ligand for members of the frizzled family of seven transmembrane receptors and is able to inhibit Wnt/β-catenin signaling as well as activate alternative non-beta-catenin dependent signaling pathways. WNT5A induces axon branching in cultured sympathetic neurons. Homozygous mice are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues.

When bred to B6.Cg-Tg(Wnt1-cre)11Rth Tg(Wnt1-GAL4)11Rth/J mice (Stock No. 009107), expressing cre in pluripotent neural crest cells and glial cells in the peripheral nervous system starting at E8.5, resulting offspring exhibit craniofacial abnormalities and died within a few hours after birth. They also have defects in the extension and branching of sympathetic fibers within the salivary glands, thymus, heart, spleen, and bladder starting at E16.5.

Development

A targeting vector was designed to insert a loxP site upstream of exon 2 followed by a second loxP site followed by a frt-flanked neomycin resistance (neo) cassette downstream of exon 2 of the wingless-type MMTV integration site family, member 5A (Wnt5a) gene. The construct was electroporated into 129S6/SvEvTac-derived MC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric males were bred to C57BL/6 females. Offspring were bred with 129S4/SvJaeSor-Gt(ROSA)26Sor^tm1(FLP1)Dym/J transgenic mice (Stock No. 003946) to delete the neo cassette, and progeny were crossed to remove the Flp-expressing transgene. The donating investigator reported that Wnt5a^fl/fl mice were bred to C57BL/6 mice for at least 7 generations (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Nine markers throughout the genome were found to be segregating with 129, suggesting an incomplete backcross.

Allele

Symbol: Wnt5a^tm1.1Krvl
Name: targeted mutation 1.1, Rejji Kuruvilla
MGI accession ID: MGI:5605689
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Wnt5a
Marker name: wingless-type MMTV integration site family, member 5A
Chromosome: 14
Strain of origin: 129S6/SvEvTac
Molecular note: A targeting vector was designed to insert a loxP site upstream of exon 2 followed by a second loxP site followed by a frt-flanked neomycin resistance (neo) cassette downstream of exon 2 of the gene. Flp-mediated recombination removed the neo cassette leaving exon 2 floxed.