This Card9 knockout strain is useful in studies of innate immune response to intracellular pathogens and fungal infection, as well as apoptosis.
Xin Lin, The University of Texas MD Anderson Cancer Center
The targeted Card9 gene encodes an adapter protein that contains a caspase recruitment domain (CARD), which interacts with Bcl10 and MALT1. Mutations in this gene have been associated with familial candidiasis and chronic mucocutaneous candidiasis.
These knock-out mice carry an allele in which a floxed NEO cassette (in antisense orientation) replaced exon 2, which includes the ATG start codon and encodes a partial CARD motif. Mice that are homozygous for the targeted mutation are viable and fertile.
No gene product (protein) is detected by Western blot analysis of peritoneal macrophages, thymus, lung, spleen, and lymph node tissue from homozgyotes.
After stimulation with Candida albicans, macrophages from homozygous animals produce a reduced level of proinflammatory cytokines (IL6, TNF) when compared to wildtype controls. Homozygotes exhibit increased susceptibility and sensitivity to Candida albicans, as well as Listeria monocytogenes bacterial infection and are defective in clearing the fungi and intracellular bacteria. CFA-induced Th17 differentiation is impaired in homozygous animals.
A targeting vector containing a loxP flanked NEO cassette (in antisense orientation) was used to disrupt exon 2 (including the ATG codon and coding region for the CARD domain) of the Card9 gene. The construct was electroporated into unspecified 129/Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts.
The resulting male animals were crossed to C57BL/6 female mice to achieve germline transmission. The mice were backcrossed to C57BL/6J for 10 generations
Heterozygotes were intercrossed to generate homozygotes.
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).
|Allele Name||targeted mutation 1, Xin Lin|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Card9, caspase recruitment domain family, member 9|
|Strain of Origin||129/Sv|
|Molecular Note||Most of exon 1 was replaced with a neomycin resistance cassette in antisense orientation. Absence of transcript and protein were confirmed by RT-PCR and Western blot.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Card9 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #028652 in your Materials and Methods section.