Homozygous targeted mutant mice die between implantation and embryonic day 12.5. There is great variability in phenotype, with some pups showing severe malformations, while others develop almost normally but arrest in development prior to their death. Virtually every organ of mutant embryos could be affected, resulting in dramatically smaller embryos and advanced stages of necrosis. Expression of the targeted gene has been assayed by Northern blot analysis of embryonic stem (ES) cells carrying the mutation and aberrant RNA was found; this is a loss of function mutation. These animals are useful in studies of development.
A targeting vector was designed to replace exons 2-4 with a loxP-flanked neomycin resistance (pgk-neo) cassette. This mutation introduced a stop codon before the coiled-coil region (CR1) region of the gene. The vector was electroporated into (129X1/SvJ x 129S1/Sv)F1- Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. Chimeric pups were identified and crossed with C57BL/6 females. In order to remove the floxed pgk-neo cassette, heterozygous mutant females were crossed to a CMV-cre male on a mixed C57BL/6-129SV background. The line was then maintained in the heterozygous condition on a mixed background by the donating laboratory by breeding heterozygous males of confirmed fertility with C57BL/6 and 129S2/SvPasCrl females (alternate generations).
|Allele Name||targeted mutation 1, Roel Nusse|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||mp66alpha null; p66 KO|
|Gene Symbol and Name||Gatad2a, GATA zinc finger domain containing 2A|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|Molecular Note||A targeting vector was designed to replace exons 2-4 with a loxP-flanked neomycin resistance (pgk-neo) cassette. This mutation introduced a stop codon before the coiled-coil region (CR1) region of the gene. In order to remove the floxed pgk-neo cassette, heterozygous mutant females were crossed to a CMV-cre male on a mixed C57BL/6-129SV background.|
|Mutations Made By|| |
Roel Nusse, Stanford University
When maintained as a live colony, heterozygotes may be bred. Homozygotes are embryonic lethal. Because of fertility problems in male heterozygotes, males must be tested for fertility. Female heterozygotes do not appear to have fertility problems.
When using the mp66α KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #008105 in your Materials and Methods section.