Homozygous Rad9atm1Lieb knock-out mice exhibit embryonic lethality with increased apoptosis and reduced cellular proliferation.
Howard B. Lieberman, Columbia University
Mice that are heterozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Mouse embryo fibroblasts (MEFs) cannot be derived from homozygous embryos. Homozygous null mice have an embryonic lethal phenotype, failing to develop somewhere between embryonic days 9.5 and 12.5. Homozygous mutant embryonic day 8.5 and 9.5 embryos exhibit increased apoptosis and reduced cellular proliferation. This mutant mouse strain may be useful in studies of development, DNA damage and repair, and genomic stability.
A targeting vector containing a herpes simplex virus thymidine kinase gene and an FRT-flanked neomycin resistance gene was utilized in the construction of this mutant. loxP sites were placed flanking exons 1 and 2 and the FRT-flanked neomycin selection cassette. The construct was electroporated into 129S6/SvEvTac derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to mice on an outbred background that express Cre recombinase under the control of the cytomegalovirus promoter to remove exons 1 and 2 and the neomycin selection cassette.
|Allele Name||targeted mutation 1, Howard B Lieberman|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Rad9a, RAD9 checkpoint clamp component A|
|Strain of Origin||129S/SvEv|
|Molecular Note||Floxed exons 1 and 2 were excised following upon mating with mice ubiquitously expressing Cre recombinase. PCR confirmed recombination in ES cells and mice and Western blot indicated no protein was produced in mutant ES cells.|
|Mutations Made By|| |
Howard Lieberman, Columbia University
When maintaining a live colony, these mice are bred as heterozygotes. Homozygotes have an embryonic lethal phenotype and do not survive past embryonic day 12.5.
When using the Mrad9- mouse strain in a publication, please cite the originating article(s) and include JAX stock #006085 in your Materials and Methods section.