B6;129S4-Pou5f1^tm1Jae/J

Stock number: 008204
Research areas: Developmental Biology Research, Research Tools

Description

These Oct4-neo mutant mice may be useful for the selection of induced pluripotent stem (iPS) cells.

Detailed description

Mice heterozygous for this Oct4-neo mutation are viable and fertile, harboring an IRES-GFPneo fusion cassette downstream of exon 5 of the Oct4 (Pou5f1) gene. The donating investigator reports (and attempts at The Jackson Laboratory confirm) no living homozygous animals are born. While sufficient neomycin-resistance activity is observed in embryonic stem cells, no visible GFP fluorescence is reported. When treated with specific transcription factors (Oct4, Sox2, c-Myc and Klf4), some Oct4-neo murine embryonic fibroblasts (MEFs) have the properties of induced pluripotent stem (iPS) cells. Such iPS cells have the DNA methylation, gene expression and chromatin state of embryonic stem cells and can form viable chimeras, contribute to the germ line, and generate live late-term embryos when injected into tetraploid blastocysts. These Oct4-neo mutant mice may be useful for the selection of iPS cells (i.e. epigenetic reprogramming of somatic cells into pluripotent embryonic stem cells).

Of note, these Oct4-neo mutant mice may also be used in conjunction with Oct4-EGFP mice (Stock No. 008214); a similar mutant strain bearing an EGFP-expressing mutation from the Oct4 locus.

Development

The Oct4-IRES-GFPneo targeting vector was designed to insert an IRES-GFPneo fusion cassette downstream of exon 5 of the Oct4 (Pou5f1) gene. This construct was microinjected into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells (with sufficient neomycin-resistance activity but no visible GFP fluorescence) were injected into blastocysts. Chimeric mice were bred to C57BL/6NTac mice to establish the mutant colony. The resulting Oct4-neo mutant mice were maintained on a mixed B6;129S4 genetic background prior to arrival at The Jackson Laboratory.

Allele

Symbol: Pou5f1^tm1Jae
Name: targeted mutation 1, Rudolf Jaenisch
MGI accession ID: MGI:3771368
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Pou5f1
Marker name: POU domain, class 5, transcription factor 1
Chromosome: 17
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Expressed genes: neo,neomycin resistance,bacterial
Site of expression: Embryonic stem cells.
Molecular note: The Oct4-IRES-GFPneo targeting vector was designed to insert an IRES-GFPneo fusion cassette downstream of exon 5. While sufficient neomycin-resistance activity is observed in embryonic stem cells, no visible GFP fluorescence is reported.
General note: When treated with specific transcription factors (Oct4, Sox2, c-Myc and Klf4), some Oct4-neo murine embryonic fibroblasts (MEFs) have the properties of induced pluripotent stem (iPS) cells. Such iPS cells have the DNA methylation, gene expression and chromatin state of embryonic stem cells and can form viable chimeras, contribute to the germ line, and generate live late-term embryos when injected into tetraploid blastocysts.