Fumarate hydratase conditional knockout mouse

Gene/Protein Targets

Fumarate Hydratase 1

Relevance: Germline mutations in the fumarate hydratase tumour suppressor gene predispose to leiomyomatosis, renal cysts, and renal cell cancer (HLRCC). Fh1-deficient mice develop renal cysts that have an activated hypoxia pathway, with overexpression of Hif1alpha and Hif2alpha. Hif targets, such as Glut1 and Vegf were also upregulated. Therefore the mouse may be useful for testing therapeutic interventions that target angiogenesis and HIF-prolyl hydroxylation.
Model Type: Conditional, Knockout Models
Zygosity: Homozygous
Phenotype Keywords:
Disease Keywords:
Production Details:

Following generation of the targeting construct, linearization, and electroporation into 129Sv/J ES cells, stable integrants were selected in 0.2 mg/ml Geneticin G418 medium. Homologous recombinants were identified initially by PCR and subsequently by Southern blot analysis. Targeted ES cells were injected into C57/BL6 blastocysts. ES cell clones were screened for 50 and 30 site-specific genomic integration of the targeting construct using a PCR-based assay and LATaq (Takara). PCR genotypes to assay Flp- and Cre-mediated recombination, and genotyping of eFlp and Cre expression mice were carried out using standard conditions. For the genotyping of Fh1 to distinguish between wild-type, null, and floxed alleles, a common forward primer (50-GCTCAGTCACCCATCCAAAT-30 ) and differential reverse primers (50-ACCCTGCTAGGTGTCACCAC-30 and 50-CCTGGCACTGCAGACTACAA-30 ) were used.

References:

Pollard PJ, et al., 2007 Cancer Cell. 4:311-9. PMID: 17418408 Read more
Bardella et al. 2011 J Pathol. PMID: 21630274 Read more

Notes:

Synonym: Fumarase
Conditional Description: LoxP flanking exons 2 and 3 of Fh1.
Growth/Phenotype: Fh1-/- mice died in early embryogenesis. Fh1 fl/fl with Ksp1.3/Cre are healthy until ~8 months when polyuric renal failure occurs. Postmortem shows macroscopic renal cysts.
Disease Keywords: Renal cyst, HLRCC