These mice possess loxP sites flanking exons 4-6 of the COP9 (constitutive photomorphogenic) homolog, subunit 8 (Arabidopsis thaliana)), Cops8, gene and have applications in studies of T cell activation and proliferation.
Ning Wei, Yale University
These mice possess loxP sites on either side of exons 4-6 of the targeted gene. Mice that are homozygous for this allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 4-6 deleted in the cre-expressing tissue(s).
When bred to a strain with Cre recombinase expression in T cells, this mutant mouse strain may be useful in studies of T cell activation and proliferation.
A targeting vector containing a FRT site flanked PGK-NEO cassette and a loxP site was utilized in the construction of this mutant. This selection cassette was inserted upstream of exon 4 of the targeted gene, and another loxP site was inserted downstream of exon 6. This construct was electroporated into 129S1/Sv-Oca2 + Tyr + Kitl + derived W9.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were backcrossed to C57BL/6 for 4 or 5 generations. To remove the selection cassette, the mice were crossed to transgenic mice (backcrossed 3 to 4 times to the C57BL/6 genetic background) expressing Flpe recombinase under the control of the beta-actin promoter. The donating investigator reported that mice that no longer retained the cassette but did retain the loxP-flanked exons 4-6 were backcrossed to C57BL/6J (see SNP note below) for 4 generations. The mice no longer carry the Flpe recombinase transgene. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 2 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
|Allele Name||targeted mutation 1, Ning Wei|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Cops8, COP9 signalosome subunit 8|
|Strain of Origin||129S1/Sv-Oca2+ Tyr+ Kitl+|
|Molecular Note||An FRT flanked neo cassette with a 3' loxP site was inserted between exon 3 and 4, and an additional loxP site was inserted between exons 6 and 7. Flp-mediate recombination to remove the neo cassette was accomplished using Tg(ACTFLPe)9205Dym. Insertion was confirmed by Southern blot analysis and PCR genotyping.|
|Mutations Made By|| |
Ning Wei, Yale University
When maintaining a live colony, these mice can be bred as homozygotes.
When using the B6.129S1(Cg)-Cops8tm1Nwe/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #017444 in your Materials and Methods section.