Mice that are homozygous for the targeted mutation are viable and fertile. These mice carry the "C282Y" mutant Hfe allele, which encodes a tyrosine to cysteine amino acid substitution at codon 282. Expression of the missense mutation is detected by RT-PCR analysis. Homozygotes exhibit hemochromatosis, accumulating hepatic iron postnatally. These mice have a phenotype that is intermediate between wildtype mice and mice that are homozygous for the null allele, Hfetm2Nca. This mutant mouse strain may be useful in studies of hereditary hemochromatosis.
A targeting vector containing the C282Y missense mutation, a tyrosine to cysteine substitution at codon 282, and neomycin resistance and cytosine deaminase genes was used to insert the missense mutation into the endogenous locus. The construct was electroporated into 129S6/SvEvTac derived TC-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts.
|Allele Name||targeted mutation 1.1, Nancy C Andrews|
|Allele Type||Targeted (Hypomorph, Humanized sequence)|
|Allele Synonym(s)||(Y); C282Y; Hfe845A|
|Gene Symbol and Name||Hfe, hemochromatosis|
|Promoter||Hfe, hemochromatosis, mouse, laboratory|
|Strain of Origin||129S6/SvEvTac|
|Molecular Note||This hypomorphic allele was generated by targeting exon 4 with a c.881G>A substitution, which causes a cysteine to tyrosine missense mutation at codon 294 (p.C294Y). This mutation is the equivalent of the p.C282Y mutation found in human hereditary hemochromatosis (HE) patients. The substitution disrupts an intramolecular disulfide bond and putatively affects Beta2-microglobulin binding. RT-PCR analysis confirmed the presence of properly spliced transcript in mutant mice. The loxP site flanked neomycin resistance gene cassette that was inserted into intron 4 was removed through cre-mediated recombination.|
|Mutations Made By|| |
Nancy Andrews, Duke University School of Medicine
The resulting chimeric animals were crossed to129S6/SvEvTac mice, and then crossed to a 129 adenovirus EIIa promoter driven cre deleter strain to remove the floxed selection cassette.
When using the C282Y mouse strain in a publication, please cite the originating article(s) and include JAX stock #005063 in your Materials and Methods section.