B6;129S4-Pou5f1^tm2Jae/J

Stock number: 008214
Product name: Oct4-GFP
Research areas: Research Tools

Description

Mice homozygous for this Oct4-EGFP mutation harbor an IRES-EGFP fusion cassette downstream of the stop codon of the Oct4 (Pou5f1) gene. These mutant mice may be useful for the selection of induced pluripotent stem (iPS) cells, or more generally for fluorescent labeling of embryonic stem cells.

Detailed description

Mice homozygous for this Oct4-EGFP mutation are viable and fertile. They harbor an IRES-EGFP fusion cassette downstream of the stop codon of the Oct4 (Pou5f1) gene. When treated with specific transcription factors (Oct4, Sox2, c-Myc and Klf4), some Oct4-EGFP 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-EGFP mutant mice may be useful for fluorescent labeling of embryonic stem cells, as well as for the selection of iPS cells (i.e. epigenetic reprogramming of somatic cells into pluripotent embryonic stem cells).

Of note, these Oct4-EGFP mutant mice may also be used in conjunction with Oct4-neo mice (Stock No. 008204); a similar mutant strain bearing a neo selection cassette in the Oct4 locus.

Development

A targeting vector was designed to insert an "IRES-EGFP-floxed NEO" cassette (containing an internal ribosomal entry site (IRES), enhanced green fluorescent protein (EGFP) sequence, and a loxP-flanked neo) between the stop codon and endogenous polyA signal of the targeted gene. This construct was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre expressing plasmid to remove the neo selection cassette. Chimeric mice were established. The resulting Oct4-EGFP mutant mice were maintained on a mixed B6;129S4 genetic background prior to arrival at The Jackson Laboratory.

Allele

Symbol: Pou5f1^tm2Jae
Name: targeted mutation 2, Rudolf Jaenisch
MGI accession ID: MGI:3772191
Generation method: Targeted
Attributes: Reporter
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: GFP,Green Fluorescent Protein,
Site of expression: Embryonic stem cells express EGFP.
Molecular note: A targeting vector was designed to insert an "IRES-EGFP-floxed NEO" cassette (containing an internal ribosomal entry site (IRES), enhanced green fluorescent protein (EGFP) sequence, and a loxP-flanked neo) between the stop codon and endogenous polyA signal of the targeted gene. This construct was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre expressing plasmid to remove the neo selection cassette.
General note: When treated with specific transcription factors (Oct4, Sox2, c-Myc and Klf4), some Oct4-EGFP 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.