These Oct4flox mutant mice possess loxP sites flanking the promoter and exon 1 of the Pou5f1 gene. This strain may be useful for generating conditional mutations in applications related to embryonic development and the regulation pluripotency in embryonic stem cells.
Hans R. Scholer, Max Planck Institute for Molecular Biomedicine
The targeted Pou5f1 gene, also called Oct4, encodes a transcription factor containing a POU homeodomain that is important in embryonic development and embryonic stem cell pluripotency.
These Oct4flox mice possess loxP sites on either side of the promoter and exon 1 of the targeted gene. 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 the promoter and exon 1 deleted in the cre-expressing tissues.
Removal of the floxed sequence creates a null allele.
When bred to a strain with germline Cre recombinase expression (see Stock No. 008569 for example), this mutant mouse strain may be useful in studies of primordial germ cell survival.
A targeting vector containing a loxP site flanked neo selection cassette was utilized in the construction of this mutant. This selection cassette was inserted downstream of exon 1 of the targeted gene, and another loxP site was inserted upstream of exon 1 and the promoter. This construct was electroporated into 129P2/OlaHsd derived E14 embryonic stem (ES) cells which were transiently transfected with a Cre recombinase vector to remove the selection cassette. ES cells in which Cre recombination resulted in exon 1 and the promoter being flanked by loxP sites were injected into C57BL/6 blastocysts. The resulting chimeric male animals were crossed to C57BL/6 female mice. 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, Hans R Shoeler|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Pou5f1, POU domain, class 5, transcription factor 1|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||One loxP site was inserted upstream of exon1 and the essential promoter. A floxed neo cassette was inserted into intron 1. The selection cassette was excised by subsequent transient transcription of Cre recombinase in the ES cells. Northern blot analysis demonstrated that a normal sized transcript was expressed from this allele.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Oct4flox mouse strain in a publication, please cite the originating article(s) and include JAX stock #029368 in your Materials and Methods section.