Pkn1-floxed mice possess loxP sites flanking exons 13-22 of the protein kinase N1 gene. These mice may be useful in studying polarized vesicle transport.
Dianqing Wu, Yale University School of Medicine
Pkn1-floxed mice possess loxP sites flanking exons 13-22 of the protein kinase N1 (Pkn1) gene. PKN1 is a PKC-related serine/threonine-protein kinase involved in the regulation of the intermediate filaments of the actin cytoskeleton, vesicle trafficking, cell migration, tumor cell invasion and transcription regulation. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 13-21 deleted in cre-expressing tissues.
A targeting vector was designed to insert a loxP site upstream of exon 13 and a frt-flanked neomycin resistance (neo) cassette, followed by a second loxP site, downstream of exon 22, including the STOP codon, of the protein kinase N1 (Pkn1) gene. The construct was electroporated into 129 embryonic stem (ES) cells and correctly targeted ES cells were injected into blastocysts. Chimeric males were bred with C57BL/6 females and their offspring were bred with B6.Cg-Tg(ACTFLPe)9205Dym/J mice (Stock No. 005703) to delete the neo cassette. Resulting Pkn1-floxed mice were crossed to remove the Flp-expressing allele, and they were subsequently bred to C57BL/6NHsd mice for at least 5 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).
|Allele Name||targeted mutation 1.1, Dianqing Wu|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Pkn1, protein kinase N1|
|Strain of Origin||129|
|Molecular Note||The targeting vector is designed to insert a loxP site upstream of exon 13 and an FRT-flanked neomycin resistance (neo) cassette, followed by a second loxP site, downstream of exon 22, which includes the STOP codon. Flp-mediated recombination removed the FRT-flanked neo cassette leaving exons 13 through 22 floxed.|
When maintaining a live colony, mice homozygous for the floxed allele may be bred together.
When using the PKN floxed mouse strain in a publication, please cite the originating article(s) and include JAX stock #029392 in your Materials and Methods section.