C57BL/6-Tg(UBC-Pou5f1,-EGFP)18Dann/J

Stock number: 029084
Product name: cOct4
Research areas: Neurobiology Research, Research Tools

Description

When crossed to a Cre recombinase expressing strain, these cOct4 transgenic mice express exogenous Pou5f1 and EGFP in the cre-expressing tissues/cells. This strain may be useful for generating conditional mutations in lineage tracking studies of POU5F1.

Detailed description

The transcription factor encoded by the Pou5f1 gene is critical in stem cell pluripotency and embryonic development. POU5F1 is also expressed in spermatogonial stem cells and undifferentiated spermatogonia. These cOct4 transgenic mice harbor a floxed STOP cassette, mouse Pou5f1 coding sequence, a myc tag, T2A/furin ribosomal skip sequence, and EGFP coding sequence all preceded by a human ubiquitin C, UBC, gene promoter.

When the floxed stop cassette is removed via Cre recombinase expression, transgenic Pou5f1 and EGFP are expressed in the cre-expressing tissues/cells. EGFP fluorescence cannot be detected directly.

The Donating Investigator reports that mice homozygous for the transgene are viable and fertile, but produce small litters.

When bred to a strain with Cre recombinase expression in embryonic germ cells (see Stock No. 006954 for example), this mutant mouse strain may be useful in studies of spermatogenesis.

Development

A transgenic construct containing (5’ to 3’) a human ubiquitin C, UBC, gene promoter, a floxed STOP cassette (3 repeats of SV40 polyA stop sequence flanked by loxP sites), the mouse Pou5f1 coding sequence with a myc tag, T2A/furin ribosomal skip sequence, EGFP coding sequence, and BGH poly(A) sequence, was injected into the pronuclei of C57BL/6 embryos. Founder line 18 was subsequently established

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).

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. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Tg(UBC-Pou5f1,-EGFP)18Dann
Name: transgene insertion 18, Christina Dann
MGI accession ID: MGI:5790657
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Tg(UBC-Pou5f1,-EGFP)18Dann
Marker name: transgene insertion 18, Christina Dann
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: Pou5f1,POU domain, class 5, transcription factor 1,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: When the floxed stop cassette is removed via Cre recombinase expression, transgenic Pou5f1 and EGFP are expressed in the cre-expressing tissues/cells.
Molecular note: The transgenic construct contains (5' to 3') a human ubiquitin C (UB) gene promoter, a floxed STOP cassette (3 repeats of SV40 polyA stop sequence flanked by loxP sites), the mouse Pou5f1 coding sequence with a myc tag, T2A/furin ribosomal skip sequence, EGFP coding sequence, and a BGH poly(A) sequence.