Panx1fl/fl floxed mice possess loxP sites flanking exon 3-4 of the pannexin 1 (Panx1) gene. PANX1 encodes a member of the innexin family of membrane channel-forming proteins expressed at postsynaptic sites in hippocampal neurons and astrocytes in central nerve system (CNS). PANX1 forms both gap junction and hemichannels, the latter being a predominant form in most cell types. Panx1 hemichannels allow communication between intra- and extracellular compartments and serves as a diffusional pathway for ions and small molecules and metabolites. PANX1 has been implicated in neurotoxicity caused by neuronal ischemia mechanical stress. Homozygotes are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 3-4 deleted in cre-expressing tissues
For example, when crossed to FVB/N-Tg(Thy1-cre)1Vln/J mice (Stock No. 006143) expressing Cre recombinase in neurons of the postnatal cortex and hippocampus, resulting mice have increased retinal ganglion cell and neuronal cell survival, with decreased apoptosis, following ischemic injury compared to wildtype mice.
When crossed to a strain with wide spread Cre recombinase expression (see Stock No. 006054), resulting offspring exhibit behavioral alterations, enhancing anxiety and impairing object recognition and spatial learning.
A targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette upstream of exon 3, and a single loxP site downstream of exon 4 of the pannexin 1 (Panx1) gene. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and the donating investigator reports that the resulting chimeric mice were bred with to C57BL/6J mice for at least 7 generations (see SNP note below). Upon arrival, Panx1fl/fl mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony. This strain retains the Casp4del mutation present in the R1 ES cells used for the development of the strain. Casp resides on Chromosome 9, 2cM away from the Panx1 gene.
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. Five markers throughout the genome are of 129 origin, suggesting an incomplete backcross.
|Allele Name||targeted mutation 1, Valery I Shestopalov|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Panx1, pannexin 1|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|Molecular Note||A floxed neo cassette was inserted upstream of exon 3. An additional loxP site was inserted downstream of exon 4.|
|Gene Symbol and Name||Casp4, caspase 4, apoptosis-related cysteine peptidase|
|Strain of Origin||129P3/J and 129S1/SvImJ and 129S2/SvPas and 129S6/SvEvTac and 129X1/SvJ|
|Molecular Note||RT-PCR confirmed that five 129 substrains (129X1/SvJ, 129S1/SvImJ, 129S2/SvPas, 129S6/SvEvTac and 129P3/J) express a transcript that lacks exon 7 (delta110 isoform). Sequencing identified a 5 bp deletion in exon 7 that results in the fusion of exon 6 and 8, a frame-shift after proline 304 and a stop codon after 5 aberrant amino acids. This deletion is not present in C57BL/6. Western blot analysis confirmed the absence of protein expression in LPS-primed macrophage.|
When maintaining a live colony, homozygotes may be bred together.
When using the B6;129-Casp4del Panx1tm1Vshe/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #026021 in your Materials and Methods section.
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