These Cenpe conditional knock-out mice possess loxP sites flanking a nucleotide binding consensus sequence of the centromere protein E gene and may be useful for studying chromosomal instability.
Dr. Don Cleveland, Ludwig Institute for Cancer Research (UCSD)
The conditionally targeted Cenpe (centromere protein E) gene encodes a kinesin-like motor protein that is required for stabilization of microtubule capture at kinetochores during prometaphase. These conditional knock-out mice possess loxP sites flanking a nucleotide binding consensus sequence (exon P). Mice that are homozygous for the targeted mutation are viable. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon P deleted in cre-expressing tissues. Removal of the floxed sequence creates a null allele. These mice may be useful for studying chromosomal instability.
Deletion of CENPE in embryos results in developmental arrest. No embryos are observed by 7.5/8.5 dpc.
Tail vein administration of an adenovirus encoding Cre recombinase, followed by chemically induced liver injury results in a 90% incidence of misaligned chromosomes and abnormal chromosome position in anaphase in hepatocytes, however, liver function is recovered.
The targeting vector contains a loxP-flanked neomycin resistance cassette upstream of a 120 nucleotide exon, referred to as exon P, and a loxP site downstream of exon P. Exon P encodes a nucleotide binding consensus sequence (P loop); deletion of exon P creates a frameshift mutation that results in a premature stop codon at aa 82. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to C57BL/6N. At some point the neomycin cassette was removed using an unknown Cre recombinase on an unspecified background. The colony has been backcrossed to C57BL/6N for 20 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 2.1, Don W Cleveland|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||CENP-E conditional|
|Gene Symbol and Name||Cenpe, centromere protein E|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|General Note||The version of this allele that contains the neomycin selection cassette, Cenpetm2Dwc, has been analyzed.|
|Molecular Note||A 120 bp exon of the gene was flanked with a loxP site and a floxed PGK-neo cassette. Cre-mediated recombination removed the floxed neo cassette.|
When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6NJ inbred mice (Stock No. 005304).
When using the CENP-E conditional mouse strain in a publication, please cite the originating article(s) and include JAX stock #032545 in your Materials and Methods section.