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.
Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Pou5f1 | POU domain, class 5, transcription factor 1 |
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.
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.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | Embryonic stem cells express EGFP. |
Allele Name | targeted mutation 2, Rudolf Jaenisch |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Oct4-EGFP; Oct4-GFP; Oct4-iresGFP; Oct4-IRES-GFP; Pou5f1tm2(EGFP)Jae |
Gene Symbol and Name | Pou5f1, POU domain, class 5, transcription factor 1 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Embryonic stem cells express EGFP. |
Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 |
Chromosome | 17 |
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. |
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. |
Mutations Made By | Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology |
When maintaining a live colony, homozygous mice may be bred.
When using the Oct4-GFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #008214 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Pou5f1<tm2Jae> |
Frozen Mouse Embryo | B6;129S4-Pou5f1<tm2Jae>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S4-Pou5f1<tm2Jae>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S4-Pou5f1<tm2Jae>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129S4-Pou5f1<tm2Jae>/J Frozen Embryo | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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