Overview

Also Known As:Oct-4/rtTA (R26-M2rtTA;Col1a1-tetO-Oct4)

These "Oct-4/rtTA" mutant mice (R26-M2rtTA;Col1a1-tetO-Oct4) may be useful for studies of tumorigenesis and pluripotent stem cells.

Donating Investigator

Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}*

Allele Type	Gene Symbol	Gene Name
Targeted (Transactivator)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

Col1a1^{tm2(tetO-Pou5f1)Jae}

Allele Type	Gene Symbol	Gene Name
Targeted (Inducible, Inserted expressed sequence, RMCE-ready)	Col1a1	collagen, type I, alpha 1

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Research Applications

Research Tools
Cancer Research
Cell Biology Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

Mice heterozygous for both targeted mutations (R26-rtTA and Cola1a::tetO-Oct4) are viable and fertile. These "Oct-4/rtTA" mice express rtTA-M2, an optimized form of reverse tetracycline-controlled transactivator (rtTA) protein, in multiple tissues. In the absence of the tetracycline analog doxycycline (dox), Oct4 (Pou5f1) expression from the Col1a1 locus is not detected. Following dox administration, high Oct4 expression is induced in liver, bone marrow, stomach, intestine, and skin, with lower levels in the heart, lungs, kidney, spleen, and thymus; no expression was detected in the brain and testes. Dox-inducd activation of Oct4 results in dysplasia in epithelial tissues. These mutant mice may be useful for studies of tumorigenesis and pluripotent cells.

Development

The R26-M2rtTA targeted mutation has an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) followed by a β-globin intron, polyA signal and PGK-puromycin cassette, all inserted downstream of the Gt(ROSA)26Sor promoter. The targeting vector was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. In addition, these ES cells were retargeted to insert an frt-flanked PGK-Neo into the 3' UTR of the Col1a1 locus. Correctly targeted ES cells were selected and the frt-flanked PGK-Neo in the 3' UTR of the Col1a1 locus was replaced by injecting both a "flip-in plasmid" containing a splice acceptor-double polyA sequence and the tetracycline responsive element (TRE or tetO) upstream of the mouse Oct4 (Pou5f1) cDNA sequence, as well as a Flpe-expressing plasmid to facilitate tetO-Oct4 recombination into the 3' UTR of the Col1a1 locus. The resulting ES cells (now targeted with rtTA-M2 in the Gt(ROSA)26Sor locus and tetO-Oct4 in the Col1a1 locus) were injected into B6D2F1 tetraploid blastocysts. Chimeric mice were bred together for many generations prior to arrival at The Jackson Laboratory Repository.

Expression Data

Expressed Gene	rtTA, reverse tetracycline-controlled transactivator, E. coli
Site of Expression	Expresses an optimized rtTA protein (rtTA-M2). Inducible target gene expression is detected in liver, bone marrow, stomach, intestine, and skin, with lower levels in the heart, lungs, kidney, spleen, and thymus; no expression is detected in the brain and testes.
Expressed Gene	Pou5f1, POU domain, class 5, transcription factor 1, mouse, laboratory
Site of Expression	With doxycycline treatment, Pou5f1 expression from the Col1a1 locus is widespread with high levels in skin, intestine and stomach, while lower levels are seen in the heart, spleen, thymus and several other organs.

Control Suggestions

Approximate Controls

Additional Information

Considerations for Choosing Controls

Selected References

Hochedlinger K; Yamada Y; Beard C; Jaenisch R. 2005. Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. Cell 121(3):465-77PubMed: 15882627 MGI: J:98920

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Genetics

Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}*

Allele Symbol: Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}*

Allele Name	targeted mutation 1, Rudolf Jaenisch
Allele Type	Targeted (Transactivator)
Allele Synonym(s)	Gt(ROSA)26Sor^{tm1(M2rtTA)Jae}; M2; M2-rtTA; R26-M2rtTA; R26-rtTA; Rosa26-M2rtTA
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	rtTA, reverse tetracycline-controlled transactivator, E. coli
Site of Expression	Expresses an optimized rtTA protein (rtTA-M2). Inducible target gene expression is detected in liver, bone marrow, stomach, intestine, and skin, with lower levels in the heart, lungs, kidney, spleen, and thymus; no expression is detected in the brain and testes.
Strain of Origin	(C57BL/6 x 129S4/SvJae)F1
Chromosome	6
General Note	This mutation was created during the generation of mutant ES cell line KH2, which also contains a frt docking site downstream of the endogenous Col1a1 locus on Chr 11 (Col1a1). The two mutations have subsequently been bred apart.
Molecular Note	An optimized form of reverse tetracycline controlled transactivator (rtTA-M2) was inserted downstream of the Gt(ROSA)26Sor promoter and was followed by a PGK-puro selection cassette. This mutant form of rtTA termed M2 has five amino acid substitutions in the tetR moiety of tTA: S12G, E19G, A56P, D148E and H179R. This mutated form of transactivator protein has increased doxycycline sensitivity. Mice have widespread expression of the rtTA-M2 protein.
Mutations Made By	Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology

Col1a1^{tm2(tetO-Pou5f1)Jae}

Allele Symbol: Col1a1^{tm2(tetO-Pou5f1)Jae}

Allele Name	targeted mutation 2, Rudolf Jaenisch
Allele Type	Targeted (Inducible, Inserted expressed sequence, RMCE-ready)
Allele Synonym(s)	Col1a1::TetOP-Oct4; TetON-Oct4
Gene Symbol and Name	Col1a1, collagen, type I, alpha 1
Gene Synonym(s)
Promoter	tetO, tet operator,
Expressed Gene	Pou5f1, POU domain, class 5, transcription factor 1, mouse, laboratory
Site of Expression	With doxycycline treatment, Pou5f1 expression from the Col1a1 locus is widespread with high levels in skin, intestine and stomach, while lower levels are seen in the heart, spleen, thymus and several other organs.
Strain of Origin	(C57BL/6 x 129S4/SvJae)F1
Chromosome	11
Molecular Note	ES cells containing the Gt(ROSA)26Sor^{tm1(rtTA2S-M2)Jae} mutation were retargeted. Cells were injected with a "flip-in plasmid" containing a tetracycline responsive element (TRE or tetO) and mouse Oct4 (Pou5f1) cDNA sequence (and a Flpe-recombinase plasmid to facilitate tetO-Oct4 integration into the 3' UTR region of Col1a1). With doxycycline treatment, Pou5f1 expression from the Col1a1 locus is widespread with high levels in skin, intestine and stomach, while lower levels are seen in the heart, spleen, thymus and several other organs. No expression is seen in the brain or testes.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Tet Expression Systems
  - tTA/rtTA Expressing Strains
  - tTA/rtTA Responsive Strains
- Genetics Research
  - Mutagenesis and Transgenesis
  - Mutagenesis and Transgenesis: transcriptional activation
  - Mutagenesis and Transgenesis: Tetop Tet System
- Cancer Research
  - Tetop Tet System
Cancer Research
- Other
Cell Biology Research
- Transcriptional Regulation

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Col1a1^{tm2(tetO-Pou5f1)Jae}/Col1a1⁺ Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}/Gt(ROSA)26Sor⁺
involves: 129S4/SvJae C57BL/6

behavior/neurological phenotype

lethargy
- animals become lethargic with doxycycline treatment within 3-5 days

endocrine/exocrine gland phenotype

thymus atrophy
- atrophy and absence of CD4, CD8 double positive cells in doxycycline treated mice

abnormal intermediate gastric gland morphology
- hyperplastic fundic glands are seen in doxycycline treated mice

hematopoietic system phenotype

thymus atrophy
- atrophy and absence of CD4, CD8 double positive cells in doxycycline treated mice

spleen atrophy
- in doxycycline treated mice

cellular phenotype

abnormal cell proliferation
- abnormal cell proliferation is observed in several organs after 2 days of Oct4-induction
- Normal - however, complete reversion is seen with withdrawal of doxycycline treatment

homeostasis/metabolism phenotype

dehydration
- after 3-5 days of doxycycline treatment, animals display severe dehydration

immune system phenotype

thymus atrophy
- atrophy and absence of CD4, CD8 double positive cells in doxycycline treated mice

spleen atrophy
- in doxycycline treated mice

integument phenotype

abnormal epidermal layer morphology
- after 5-10 days of doxycycline treatment, mice show mild to moderate epidermal dysplasia with a decrease in differentiation in dysplastic cells

abnormal hair follicle morphology
- increased numbers of immature cells in the hair follicles of the skin are seen after 5-10 days of doxycycline

increased skin tumor incidence
- in doxycycline treated mice, tumors originating from the outer-root-sheath progenitors and invading the subcutaneous layer are seen

mortality/aging

premature death
- Normal - however, if doxycycline treatment is stopped after 5 days mice completely recover
- mice with doxycycline-induced ectopic Oct4 expression become morbid after 3-5 days of treatment and usually die after 5-10 days of treatment

neoplasm

increased skin tumor incidence
- in doxycycline treated mice, tumors originating from the outer-root-sheath progenitors and invading the subcutaneous layer are seen

digestive/alimentary phenotype

abnormal intermediate gastric gland morphology
- hyperplastic fundic glands are seen in doxycycline treated mice

abnormal small intestine morphology
- after 5 days of doxycycline treatment, proliferative zone expands; postmitotic, differentiated cells lining the villus are replaced
- the proximal part of the small intestine is most severely affected by doxycycline treatment with abnormal cells often almost obstructing the lumen
- upon cessation of treatment, cells migrate to final destinations and differentiate
- Normal - upon cessation of treatment, cells migrate to final destinations and differentiate resulting in restoration of normal morphology

abnormal forestomach morphology
- in doxycycline treated mice, cells in the forestomach show a marked atypia and increased mitotic activity

abnormal stomach epithelium morphology
- in doxycycline treated mice, forestomach epithelium is thickened and stomach shows lack of differentiation into granular and cornified cell layers compared to control mice
- the thickened epithelium consists of atypical cells with enlarged nuclei and prominent nucleoli

stomach epithelial hyperplasia
- cells show atypia and increased mitotic activity throughout the squamous epithelial layer in doxycycline treated mice

abnormal stomach glandular epithelium morphology
- after doxycycline treatment, mice display severe dysplasia and increased proliferation

abnormal stomach mucosa morphology
- pyloric mucosa contains lesions resembling high grade-dysplasia in doxycycline treated mice

abnormal intestinal epithelium morphology
- after doxycline treatment, dysplastic cells are found in the entire epithelium; cells have structural and cytological dysplasia which mimics adenocarcinoma

References

Hochedlinger K; Yamada Y; Beard C; Jaenisch R. 2005. Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. Cell 121(3):465-77PubMed: 15882627 MGI: J:98920

Additional - Col1a1^{tm2(tetO-Pou5f1)Jae} related

Additional - Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}* related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Gt(rosa)26sor Probe
- Standard PCR:Col1a1 3'UTR assay2 (flip-in)
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice that are heterozygous or homozygous for Gt(ROSA)26Sor^{tm1(rtTA*M2)Jae} and heterozygous for Col1a1^{tm2(tetO-Pou5f1)Jae} may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the Oct-4/rtTA (R26-M2rtTA;Col1a1-tetO-Oct4) mouse strain in a publication, please cite the originating article(s) and include JAX stock #006911 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

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

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Gt(ROSA)26Sor<tm1(rtTA*M2)Jae>, Heterozygous or wildtype for Col1a1<tm2(tetO-Pou5f1)Jae>

Payment Terms and Conditions

Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.

The Jackson Laboratory's Genotype Promise

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.

Terms Of Use

General Terms and Conditions

Questions about Terms of Use

Additional Use Restrictions Apply

Use of MICE by companies or for-profit entities requires a license prior to shipping.

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Also Known As:Oct-4/rtTA (R26-M2rtTA;Col1a1-tetO-Oct4)

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm1(rtTA*M2)Jae

Allele Symbol: Gt(ROSA)26Sortm1(rtTA*M2)Jae

Col1a1tm2(tetO-Pou5f1)Jae

Allele Symbol: Col1a1tm2(tetO-Pou5f1)Jae

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Col1a1tm2(tetO-Pou5f1)Jae/Col1a1+ Gt(ROSA)26Sortm1(rtTA*M2)Jae/Gt(ROSA)26Sor+involves: 129S4/SvJae * C57BL/6

behavior/neurological phenotype

endocrine/exocrine gland phenotype

hematopoietic system phenotype

cellular phenotype

homeostasis/metabolism phenotype

immune system phenotype

integument phenotype

mortality/aging

neoplasm

digestive/alimentary phenotype

References

Additional - Col1a1tm2(tetO-Pou5f1)Jae related

Additional - Gt(ROSA)26Sortm1(rtTA*M2)Jae related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

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

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}*

Allele Symbol: Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}*

Col1a1^{tm2(tetO-Pou5f1)Jae}

Allele Symbol: Col1a1^{tm2(tetO-Pou5f1)Jae}

Genotype: Col1a1^{tm2(tetO-Pou5f1)Jae}/Col1a1⁺ Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}/Gt(ROSA)26Sor⁺
involves: 129S4/SvJae C57BL/6

Additional - Col1a1^{tm2(tetO-Pou5f1)Jae} related

Additional - Gt(ROSA)26Sor^{tm1(rtTAM2)Jae}* related