STOCK Wwtr1^tm1.2Hmc Yap1^tm1.2Hmc/WranJ

Stock number: 030532
Product name: Taz flox/Yap flox
Research areas: Cancer Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

These double floxed mutant mice possess loxP sites flanking exon 2 of both the Wwtr1 and Yap1 genes. This strain may be useful for generating conditional mutations in applications related to the Hippo signaling pathway, organogenesis, tumorigenesis, and tumor suppression.

In 2024, analysis performed by The Jackson Laboratory revealed the mice we originally received, cryopreserved and have always distributed have the Wwtr1 floxed allele devoid of the frt-flanked neo and the Yap1 floxed allele devoid of the frt-flanked SA-IRES-βgeo. The absence of those sequences is expected to have no adverse effects on endogenous expression from each floxed allele nor their ability to be Cre recombined.

Detailed description

The targeted Wwtr1 gene (also known as TAZ) encodes a transcriptional coactivator that with YAP, encoded by the Yap1 gene, are downstream effectors of the Hippo pathway. YAP/WWTR1 are involved in the regulation of cell proliferation, differentiation, and survival as well as mechanotransduction, epithelial-mesenchymal transition in the extracellular matrix, organ size, and stem cell self-renewal. When YAP/WWTR1 are translocated to the nucleus (inactivated Hippo signaling), they act as coactivators for TEA domain (TEAD) transcription factors. Dysregulated Hippo signaling has been associated with a number of human cancers. Mutations in the Yap1 gene have been associated with ocular coloboma, with or without hearing impairment, cleft lip/palate, and/or mental retardation.

These double floxed mice possess loxP sites on either side of exon 2 of the targeted Wwtr1 gene and loxP sites on either side of exon 2 of the targeted Yap1 gene. Mice that are homozygous for both of these alleles are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 2 of Wwtr1 and exon 2 of Yap1 deleted in the cre-expressing tissues. Removal of the floxed sequence creates null alleles.

When bred to a strain with Cre recombinase expression in nephron progenitor cells, this mutant mouse strain may be useful in studies of kidney organogenesis.

When bred to a strain with Cre recombinase expression in Schwann cells (see Stock No. 017927 for example), this mutant mouse strain may be useful in studies of myelination and axonal sorting during neuronal development.

When bred to a strain with Cre recombinase expression in villus and crypt epithelial cells of the small and large intestines (see Stock No. 021504 for example), this mutant mouse strain may be useful in studies of intestinal adenoma and tumor suppression.

Development

For the Wwtr1 floxed allele, a targeting vector containing a FRT-site flanked NEO cassette was utilized in the construction of this mutant. This selection cassette, followed by a loxP site, was inserted upstream of exon 2, and another loxP site was inserted downstream of exon 2. This construct was electroporated into (129X1/SvJ x 129S1/Sv)F1- Kitl⁺-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. Breeding to Flpe-expressing mice removed the FRT-flanked NEO. The mice were maintained on a mixed background including ICR, C57BL/6 and 129S (and the FLPe allele was bred away).

For the Yap1 floxed allele, a targeting vector containing a FRT-site flanked Splice Acceptor-IRES-βgeo (lacZ/neo) cassette was utilized in the construction of this mutant. A loxP site followed by this selection cassette, was inserted downstream of exon 2, and another loxP site was inserted upstream of exon 2. This construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1 derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. Breeding to Flpe-expressing mice removed the FRT-flanked SA-IRES-βgeo. The mice were maintained on a mixed genetic background (and the FLPe allele was bred away).

The two floxed lines were then intercrossed to generate double homozygous floxed mice. Upon arrival in 2017, 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: Yap1^tm1.2Hmc
Name: targeted mutation 1.2, Helen McNeill
MGI accession ID: MGI:7581996
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Yap1
Marker name: yes-associated protein 1
Marker synonyms: COB1; Yap; YAP-1; YAP2; YAP65; YKI; yorkie
Chromosome: 9
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: An FRT flanked splice acceptor-IRES-betageo cassette and loxP site were inserted in intron 1 and a loxP site was inserted in intron 2 via homologous recombination. Breeding to Flpe-expressing mice removed the FRT-flanked SA-IRES-betageo.
General note: The version of this allele that contains the IRES-betageo cassette, Yap1^tm1Hmc, has been analyzed.

Allele

Symbol: Wwtr1^tm1.2Hmc
Name: targeted mutation 1.2, Helen McNeill
MGI accession ID: MGI:7581994
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Wwtr1
Marker name: WW domain containing transcription regulator 1
Marker synonyms: 2310058J06Rik; 2610021I22Rik; TAZ; transcriptional coactivator with PDZ binding motif
Chromosome: 3
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: A loxP site was inserted in intron 1 and a loxP site and FRT-flanked neo cassette were inserted in intron 2 via homologous recombination. Breeding to Flpe-expressing mice removed the FRT-flanked NEO.
General note: The version of this allele that contains the neo cassette, Wwtr1^tm1Hmc, has been analyzed.