Strain Name:

B6.129S6-Pou5f1tm1Grc/J

Stock Number:

023515

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Availability:

Cryopreserved - Ready for recovery

Use Restrictions Apply, see Terms of Use
This modified Pou5f1 (POU domain, class 5, transcription factor 1; also called Oct4) strain may be used to further study the role of histones in gene regulation and is part of a chromatin in vivo assay (CiA) system employing chemically-induced proximity to initiate and terminate chromatin modifications in living cells.

Description

The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.

Strain Information

Type Congenic; Mutant Strain; Targeted Mutation;
Additional information on Genetically Engineered and Mutant Mice.
Visit our online Nomenclature tutorial.
Additional information on Congenic nomenclature.
Specieslaboratory mouse
 
Donating Investigator Gerald R Crabtree,   Stanford University Medical Center

Description
Upon cellular differentiation, Pou5f1 (POU domain, class 5, transcription factor 1; also called Oct4), highly expressed in embryonic stem cells, is rapidly and completely silenced through a series of events including histone H3K9 methylation, heterochromatic protein 1 (HP1) binding, and DNA methylation. HP1 can form oligomers, which are thought to bridge neighboring nucleosomes and mediate chromatin condensation.

This mutant mouse strain may be used to further study the role of histones in gene regulation and is part of a chromatin in vivo assay (CiA) system employing chemically-induced proximity to initiate and terminate chromatin modifications in living cells. It enables rapid addition and removal of chromatin regulatory activities to a genetically modified Pou5f1 allele in any cell type by using small molecule-mediated recruitment. Two arrays of different DNA binding sites (12xZFHD1 and 5xGAL4) were placed upstream of the Pou5f1 promoter and an in-frame nuclear enhanced green fluorescent protein (EGFP) reporter replaces the first exon of the gene.

In an embryonic stem cell or embryonic fibroblast model, the ZFHD1 and GAL4 DNA binding domains can be fused to CIP anchor partners (e.g., FKBP12 or ABI1, respectively). Upon induction by small membrane-permeable molecules (e.g., rapamycin or plant hormone abscisic acid (ABA)), these fused domains can reversibly bind a second set of protein chimeras containing a protein of interest fused to the CIP recruitment partner (e.g., csHP1α fused to FRB or VP16 fused to PYL1).

Selective recruitment of HP1a through the addition of rapamycin has been found to induce H3K9me3 (H3K9 trimethylation) at the Pou5f1 reporter locus and cause subsequent linear spreading in cis over a distance of 10 kbp to form a heterochromatic domain with features of position effect variegation (PEV). Infection with full-length HP1a and csHP1a fusion proteins induces complete silencing of gene/GFP expression, as measured by flow cytometry. The presence of the two DNA binding domains enables temporal resolution of the dynamic processes involved with HP1α-mediated repression and facilitates studies of heterochromatin stabilization.

Mice homozygous for this mutation die before birth, heterozygous are viable and fertile. Targeted ES cells retained good morphology and provided real-time fluorescence-based readout of gene expression at single cell resolution.

Development
A BAC containing the mouse Pou5f1 (Oct4) locus was manipulated through recombineering. The construct was flanked with homology arms of approximately 3.7 kbp upstream and 10.9 kbp downstream. Two distinct DNA binding sequence arrays were added at 277 bp upstream of the transcriptional start of of the gene, 12 x ZFHD1 (TAATGATGGGCG) and 5 x Gal4 (CGGAGTACTGTCCTCCGAG). A nuclear EGFP was inserted at the ATG of exon 1, exons 2-5 were deleted after insertion to avoid expression of endogenous Pou5f1 from the reporter allele. A floxed neomycin resistance cassette was inserted behind the EGFP reporter for positive selection and thymidine kinase was inserted behind the homology arms for negative selection. Gene targeting was done in 129S6/SvEvTac-derived TC1 embryonic stem (ES) cells. This strain was backcrossed to C57BL/6 for 4 generations by the donating laboratory.

Control Information

  Control
   Wild-type from the colony
 
  Considerations for Choosing Controls

Related Strains

View Strains carrying other alleles of Pou5f1     (10 strains)

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms provided by MGI
      assigned by genotype

The following phenotype information is associated with a similar, but not exact match to this JAX® Mice strain.

Pou5f1tm1Grc/Pou5f1tm1Grc

        involves: 129S6/SvEvTac * C57BL/6
  • mortality/aging
  • complete prenatal lethality
    • mice die before birth   (MGI Ref ID J:199710)
View Research Applications

Research Applications
This mouse can be used to support research in many areas including:

Cell Biology Research
Transcriptional Regulation

Genes & Alleles

Gene & Allele Information provided by MGI

 
Allele Symbol Pou5f1tm1Grc
Allele Name targeted mutation 1, Gerald R Crabtree
Allele Type Targeted (Null/Knockout, Reporter)
Strain of Origin129S6/SvEvTac
Gene Symbol and Name Pou5f1, POU domain, class 5, transcription factor 1
Chromosome 17
Gene Common Name(s) OCT3; OCT4; OTF-3; OTF3; OTF4; Oct-3; Oct-3/4; Oct-4; Oct3/4; Otf-3; Otf-4; Otf3; Otf3-rs7; Otf3g; Otf4; octamer binding transcription factor 3; octamer binding transcription factor 3 related sequence 7; octamer binding transcription factor 4;
Molecular Note A BAC containing the mouse locus was manipulated through recombineering. The construct was flanked with homology arms of approximately 3.7 kbp upstream and 10.9 kbp downstream. Two distinct DNA binding sequence arrays were added at -277 bp upstream of the transcriptional start of of the gene, 12 x ZFHD1 (TAATGATGGGCG) and 5 x Gal4 (CGGAGTACTGTCCTCCGAG). A nuclear EGFP was inserted at the ATG of exon 1, exons 2-5 were deleted after insertion to avoid expression of endogenous Pou5f1 from the reporter allele. A floxed neomycin resistance cassette was inserted behind the EGFP reporter for positive selection and thymidine kinase was inserted behind the homology arms for negative selection. [MGI Ref ID J:199709] [MGI Ref ID J:199710]

Genotyping

Genotyping Information

Genotyping Protocols

Pou5f1tm1Grc, Standard PCR


Helpful Links

Genotyping resources and troubleshooting

References

References provided by MGI

Selected Reference(s)

Crabtree GR. 2013. Direct Data Submission for Pou5fl MGI Direct Data Submission :.  [MGI Ref ID J:199710]

Hathaway NA; Bell O; Hodges C; Miller EL; Neel DS; Crabtree GR. 2012. Dynamics and memory of heterochromatin in living cells. Cell 149(7):1447-60. [PubMed: 22704655]  [MGI Ref ID J:199709]

Health & husbandry

The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.

Health & Colony Maintenance Information

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200.

Colony Maintenance

Breeding & HusbandryHeterozygotes are viable and fertile. Homozygotes die before birth.

Pricing and Purchasing

Pricing, Supply Level & Notes, Controls


Pricing for USA, Canada and Mexico shipping destinations View International Pricing

Cryopreserved

Cryopreserved Mice - Ready for Recovery

Price (US dollars $)
Cryorecovery* $2140.00
Animals Provided

At least two mice that carry the mutation (if it is a mutant strain) will be provided. Their genotypes may not reflect those discussed in the strain description. Please inquire for possible genotypes and see additional details below.

Standard Supply

Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.

Supply Notes

  • Cryorecovery - Standard.
    Progeny testing is not required.

    The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided, their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders are needed. Animals typically ship between 10 and 14 weeks from the date of your order. If a second cryorecovery is needed in order to provide the minimum number of animals, animals will ship within 25 weeks. IMPORTANT NOTE: The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

    Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 (from U.S.A., Canada or Puerto Rico only) or 1-207-288-5845 (from any location).

Pricing for International shipping destinations View USA Canada and Mexico Pricing

Cryopreserved

Cryopreserved Mice - Ready for Recovery

Price (US dollars $)
Cryorecovery* $2782.00
Animals Provided

At least two mice that carry the mutation (if it is a mutant strain) will be provided. Their genotypes may not reflect those discussed in the strain description. Please inquire for possible genotypes and see additional details below.

Standard Supply

Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.

Supply Notes

  • Cryorecovery - Standard.
    Progeny testing is not required.

    The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided, their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders are needed. Animals typically ship between 10 and 14 weeks from the date of your order. If a second cryorecovery is needed in order to provide the minimum number of animals, animals will ship within 25 weeks. IMPORTANT NOTE: The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

    Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 (from U.S.A., Canada or Puerto Rico only) or 1-207-288-5845 (from any location).

View USA Canada and Mexico Pricing View International Pricing

Standard Supply

Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.

Control Information

  Control
   Wild-type from the colony
 
  Considerations for Choosing Controls
  Control Pricing Information for Genetically Engineered Mutant Strains.
 

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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.
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