B6.129S2-F5^tm2Dgi/J

Stock number: 004080
Research areas: Hematological Research

Description

These F5 Leiden mutation mice serve as a model for blood coagulation defects.

Detailed description

These mice carry the F5 Leiden mutation whereby the arginine at position 504 is replaced by a glutamine disrupting a cleavage site necessary for the inactivation of the factor V protein. Mice that are homozygous for this allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Clotting activity is indistinguishable from that of wildtype mice. Rarely, spontaneous vascular thrombo-embolic events are observed. Analysis of tissue fibrin content indicates increased deposition of fibrin in multiple tissues. When maintained on a mixed B6;129 genetic background, mice homozygous for this allele experience disseminated intravascular thrombosis resulting in increased fetal loss. This loss is not observed on a congenic C57BL/6J genetic background. This mutant mouse strain represents a model useful in research examining the disease aspects of the Leiden mutation and thrombosis in general.

Development

A targeting vector containing F5 exons 7-11, with a TK/neo cassette flanked by loxP sites located in intron 10 and a point mutation in exon 10 (mutation R504Q) was introduced into 129S2/SvPas-derived D3 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with supercoiled pMC-Cre plasmid DNA to remove the TK/neo cassette. ES cells bearing the R504Q mutation and bereft of the TK/Neo cassette were injected into C57BL/6J blastocysts. The resulting chimeric male animals were backcrossed to C57BL/6J females. Mutant mice were then backcrossed to C57BL/6J for twelve generations before being made homozygous.

Allele

Symbol: F5^tm2Dgi
Name: targeted mutation 2, David Ginsburg
MGI accession ID: MGI:1931528
Generation method: Targeted
Attributes: Hypomorph; Humanized sequence
Synonyms: FV^Leiden; Fv^Q; FvL; FVLq; FvR504Q
Marker symbol: F5
Marker name: coagulation factor V
Marker synonyms: Ac2-120; Cf5; Cf-5; FVL; PCCF; RPRGL1; THPH2
Chromosome: 1
Strain of origin: 129S2/SvPas
Molecular note: A point mutation was introduced which altered the codon corresponding to amino acid 532 from arginine to glutamine (p.R532Q or p.R504Q in reference to mature protein with signal peptide removed). The loxP site flanked neomycin resistance gene cassette inserted into intron 10 was removed by subsequent Cre-mediated recombination. This mutation is the equivalent of the human p.R506Q Factor 5 Leiden mutation (or UNIPROT:P12259:p.R534Q for peptide including signal peptide).