B6.129S4(Cg)-Vcl^tm1Ross/J

Stock number: 028451
Product name: Vcl^fl
Research areas: Cell Biology Research, Developmental Biology Research, Research Tools

Description

Vcl^fl/fl floxed mice possess loxP sites flanking exon 3 of the vinculin gene making this strain useful for studying the regulation of cell-cell and cell-matrix adhesion.

Detailed description

Vcl^fl/fl floxed mice possess loxP sites flanking exon 3 of the vinculin (Vcl) gene. Vinculin is a cytoskeletal protein associated with cell-cell and cell-matrix junctions, where it is involved in anchoring actin filaments to the cell membrane. Mice that are homozygous for this allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 3 deleted in the cre-expressing tissues.

When bred to mice expressing Cre Recombinase in cardiac myocytes, Vcl deletion results in a disruption of the adherens junctions in the myocardium, causing death by 3 months of age by either sudden death or dilated cardiomyopathy with progressive heart failure.

Development

A targeting vector was designed by Dr Robert Ross (University of California, San Diego School of Medicine) to insert a loxP site upstream of exon 3, and a second loxP site downstream of exon 3 of the vinculin (Vcl) gene. A frt-flanked neomycin resistance (neo) cassette was placed downstream of the 3’ loxP site. The construct was electroporated into 129S4/SvJae-derived LW1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with B6.Cg-Tg(ACTFLPe)9205Dym/J (Stock No. 005703) transgenic mice to delete the neo cassette. Progeny were crossed to remove the Flp-expressing transgene, and resulting Vcl^fl/fl mice were bred to Black Swiss mice. Subsequently, mice were bred to C57BL/6J mice for 15 generations. 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: Vcl^tm1Ross
Name: targeted mutation 1, Robert S Ross
MGI accession ID: MGI:3769142
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Vcl
Marker name: vinculin
Chromosome: 14
Strain of origin: 129S4/SvJae
Molecular note: An frt-flanked neo cassette with a 5' loxP site was inserted downstream of exon 3. An additional loxP site was inserted upstream of exon 3. Flp-mediated recombination was used to remove the neo cassette leaving a floxed exon 3.