B6.129P2(FVB)-Ptk2^tm1.1Guan/J

Stock number: 031956
Product name: FAK^flox
Research areas: Cardiovascular Research, Cell Biology Research, Developmental Biology Research, Research Tools

Description

FAK^flox mice possess loxP sites flanking exon 3 of the PTK2 protein tyrosine kinase 2 (Ptk2) gene. This strain may be useful for studying angiogenesis and vascular development in late embryogenesis.

Detailed description

These FAK^flox mutant mice possess loxP sites flanking exon 3 of the PTK2 protein tyrosine kinase 2 (Ptk2) gene. PTK2, also known as Focal Adhesion Kinase (FAK), is a cytoplasmic tyrosine kinase that plays a role in integrin-mediated signal transduction pathways that regulate cell spreading and migration, cell survival, and cell cycle progression. FAK also plays a role in vascular development. Recent studies have also revealed novel functions in the regulation of epithelial-mesenchymal transition (EMT), an important developmental program exploited by cancer cells. Exon 3 encodes the FERM domain which is required for the binding of regulatory protein partners, converting FAK into its active state. Mice that are homozygous for this allele are viable. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 3 deleted in cre-expressing tissues.

For example, when crossed to B6.Cg-Tg(Tek-cre)12Flv/J mice (Stock No. 004128) expressing Cre recombinase in endothelial cell, resulting offspring contain a frame-shift mutation because of the direct splicing from exon 2 (containing the ATG initiation codon) to exon 4, which produces a small truncated and nonfunctional peptide (~70 aa) lacking the majority of FAK sequences. These mice are embryonic lethal by E11.5 and exhibit growth retardation, abnormal embryonic and extra embryonic tissue development, and abnormal vascular development.

Development

The FAK^flox targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette, in opposite orientation, downstream of exon 3, and a single loxP site upstream of exon 3 of the PTK2 protein tyrosine kinase 2 (Ptk2) gene. The construct was electroporated into 129P2/OlaHsd-derived E14.1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with C57BL/6J mice. Subsequently, mice were bred to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J mice (Stock No. 003724) to delete the neo cassette. Resulting mice contained multiple gene rearrangements; intact floxed-exon 3, intact floxed-neo cassette, or excision of both exon 3 and the neo cassette. Mice containing only the floxed-exon 3, were bred with C57BL/6J mice for at least 7 generations and they were subsequently bred to C57BL/6Hsd mice for one generation. The donating investigator reports that the mice are 64% B6N and 36% B6J in 2017. Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Ptk2^tm1.1Guan
Name: targeted mutation 1.1, Jun-Lin Guan
MGI accession ID: MGI:3583806
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: FAK^flox
Marker symbol: Ptk2
Marker name: PTK2 protein tyrosine kinase 2
Marker synonyms: Fadk; FADK 1; FAK; FAK1; FRNK; p125FAK; pp125FAK; PPP1R71
Chromosome: 15
Strain of origin: 129P2/OlaHsd
Molecular note: A targeting vector was designed to flank exon 3 with loxP sites. Western blot confirmed loxP sequences did not interfere with normal expression of the protein in mutants.