This Card6 knockout strain may be useful in studies of signal transduction and NF-ΚB activation.
Dr. Tak Mak, University Health Network / University of Toronto
The targeted Card6 gene encodes a protein that carries a CARD, caspase recruitment domain, interacts with microtubules, modulates NF-ΚB activation and is upregulated beta IFN and gamma IFN.
These knock-out mice carry a mutation in which exon 4 has been deleted via Cre-mediated recombination. No gene product (mRNA or ΔCARD6S variant protein) is detected by RT-PCR analysis of splenocytes or Western blot analysis of bone marrow and thymocytes from homozygotes.
Mice that are homozygous for the targeted mutation are viable, fertile, and show no overt phenotype. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background.
A targeting vector containing a FRT site flanked Neo selection cassette and a 3’ end loxP 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 4. The construct was electroporated into 129P2/OlaHsd derived E14K embryonic stem (ES) cells.
Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were tested for germline transmission. Heterozygotes were crossed with Cre deleter mice carrying the Tg(CMV-cre)1Cgn/J transgene, on an unspecified genetic background, to delete the floxed exon 4.
The mice were then backcrossed to C57BL/6 for 10 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background.
Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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 43 markers throughout the genome suggested a C57BL/6 genetic background, all 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 1.1, Almut Dufner|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Card6, caspase recruitment domain family, member 6|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||The last coding exon was removed by germ-line, cre-mediated recombination. The absence of protein product was confirmed by western blot analysis on thymus and bone marrow extracts.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Card6 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #029253 in your Materials and Methods section.