These Ezh2F/F mutant mice possess loxP sites flanking exons 14-15 of the zeste homolog 2 (Ezh2) gene. This strain may be useful for studying chromatin condensation and gene silencing.
Stuart H Orkin, Harvard University
These Ezh2F/F mice possess loxP sites flanking exons 14-15 of the zeste homolog 2 (Ezh2) gene. EZH2 is part of polycomb repressive complexes (PRC) 2. PRC2 is involved in transcriptional repression by the addition of three methyl groups to histone 3 during chromatin condensation. 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 14-15 deleted in cre-expressing tissues.
For example, when crossed to B6.Cg-Tg(Mx1-cre)1Cgn/J mice (Stock No. 003556) expressing Cre recombinase after poly I:C treatment, Ezh2 deficiency leads to the development of spontaneous T-cell leukemia (T-ALL) 122-281 days following treatment.
A targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette upstream of exon 14, and a single loxP site downstream of exon 15 of the enhancer of zeste homolog 2 (Ezh2) gene. The construct was electroporated into 129S1/Sv-Oca2+ Tyr+ Kitl+-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a Cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exons 14-15, intact floxed-neo cassette, or excision of both exons and the neo cassette. Correctly targeted ES cells, containing only the floxed-exons 14-15, were injected into blastocysts and resulting chimeric mice were maintained on a mixed background. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 2, Stuart Orkin|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Ezh2, enhancer of zeste 2 polycomb repressive complex 2 subunit|
|Strain of Origin||129S1/Sv-Oca2+ Tyr+ Kitl+|
|Molecular Note||A floxed neo cassette was inserted into intron 13 and an additional loxP site was inserted into intron 15. Transient cre expression was used to remove the neo cassette in ES cells leaving exons 14 and 15 floxed.|
When maintaining a live colony mice homozygous for the floxed allele may be bred together.
When using the Ezh2fl mouse strain in a publication, please cite the originating article(s) and include JAX stock #022616 in your Materials and Methods section.