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| These "Oct-4/rtTA" mutant mice may be useful for studies of tumorigenesis and pluripotent stem cells. | |||||||||
- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
Description
Strain Information
Former Names B6;129-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm2(tetO-Pou5f1)Jae/J (Changed: 12-APR-07 ) Type Mutant Stock; Targeted Mutation; Species laboratory mouse Generation ?+N1F1 (08-DEC-07) Donating Investigator Rudolf Jaenisch, Whitehead Institute (MIT) 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
An optimized form of reverse tetracycline controlled transactivator (rtTA-M2) was introduced downstream of the Gt(ROSA)26Sor promoter by gene targeting in (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. The modified ES cells were retargeted to allow the introduction of a tetO-Oct4 transgene into the 3? UTR of the Col1a1 locus by frt/Flpase-mediated site-specific integration. Briefly, an frt site was introduced into the 3' UTR region of Col1a1. locus by gene targeting. Correctly targeted ES cells were then cotransfected with a "flip-in plasmid" containing mouse Oct4 (Pou5f1) cDNA sequence under the control of a tetracycline responsive element (TRE or tetO) and a Flpe-recombinase plasmid to facilitate tetO-Oct4 integration into the 3' UTR region of Col1a1. The resulting ES cells were injected into B6D2F1 tetraploid blastocysts. Chimeric mice were bred together for many generations prior to arrival at The Jackson Laboratory.
Control Information
| Control | ||
|---|---|---|
| 101043 B6129SF1/J | (approximate) | |
| 101045 B6129SF2/J | (approximate) | |
| Considerations for Choosing Controls | ||
Related Strains
Strains carrying Gt(ROSA)26Sortm1(rtTA*M2)Jae allele
006965 B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae/J View Strains carrying Gt(ROSA)26Sortm1(rtTA*M2)Jae (1 strain)
002495 B6;129S4-Col1a1tm1Jae/J 002197 C57BL/6-Col1a1Mov13/J
008204 B6;129S4-Pou5f1tm1Jae/J 008214 B6;129S4-Pou5f1tm2Jae/J 004654 B6;CBA-Tg(Pou5f1-EGFP)2Mnn/J
Additional Web Information
Tet Expression Systems
Phenotype
Phenotype Information
Mammalian Phenotype Terms assigned by genotype
Col1a1tm2(tetO-Pou5f1)Jae/Col1a1+ Gt(ROSA)26Sortm1(rtTA*M2)Jae/Gt(ROSA)26Sor+
involves: 129S4/SvJae * C57BL/6
- life span-post-weaning/aging
- abnormal induced morbidity/mortality (MGI Ref ID J:98920)
- mice with doxycycline-induced ectopic Oct4 expression become morbid after 3-5 days of treatment and usually die after 5-10 days of treatment
- however, if doxycycline treatment is stopped after 5 days mice completely recover
- digestive/alimentary phenotype
- abnormal intestinal epithelium morphology (MGI Ref ID J:98920)
- after doxycline treatment, dysplastic cells are found in the entire epithelium; cells have structural and cytological dysplasia which mimics adenocarcinoma
- abnormal small intestine morphology (MGI Ref ID J:98920)
- the proximal part of the small intestine is most severely affected by doxycycline treatment with abnormal cells often almost obstructing the lumen
- after 5 days of doxycycline treatment, proliferative zone expands; postmitotic, differentiated cells lining the villus are replaced
- upon cessation of treatment, cells migrate to final destinations and differentiate upon cessation of treatment, cells migrate to final destinations and differentiate resulting in restoration of normal morphology
- abnormal stomach morphology (MGI Ref ID J:98920)
- in doxycycline treated mice, cells in the forestomach show a marked atypia and increased mitotic activity
- abnormal stomach epithelium morphology (MGI Ref ID J:98920)
- 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
- abnormal stomach glandular epithelium morphology (MGI Ref ID J:98920)
- after doxycycline treatment, mice display severe dysplasia and increased proliferation
- abnormal gastric mucosa morphology (MGI Ref ID J:98920)
- pyloric mucosa contains lesions resembling high grade-dysplasia in doxycycline treated mice
- abnormal gastric gland (MGI Ref ID J:98920)
- hyperplastic fundic glands are seen in doxycycline treated mice
- stomach epithelial hyperplasia (MGI Ref ID J:98920)
- cells show atypia and increased mitotic activity throughout the squamous epithelial layer in doxycycline treated mice
- homeostasis/metabolism phenotype
- dehydration (MGI Ref ID J:98920)
- after 3-5 days of doxycycline treatment, animals display severe dehydration
- cellular phenotype
- abnormal cell proliferation (MGI Ref ID J:98920)
- abnormal cell proliferation is observed in several organs after 2 days of Oct4-induction
- however, complete reversion is seen with withdrawal of doxycycline treatment
- behavior/neurological phenotype
- lethargy (MGI Ref ID J:98920)
- animals become lethargic with doxycycline treatment within 3-5 days
- skin/coat/nails phenotype
- abnormal epidermal layer morphology (MGI Ref ID J:98920)
- 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 (MGI Ref ID J:98920)
- increased numbers of immature cells in the hair follicles of the skin are seen after 5-10 days of doxycycline
- skin tumor (MGI Ref ID J:98920)
- in doxycycline treated mice, tumors originating from the outer-root-sheath progenitors and invading the subcutaneous layer are seen
- hematopoietic system phenotype
- spleen atrophy (MGI Ref ID J:98920)
- in doxycycline treated mice
- thymus atrophy (MGI Ref ID J:98920)
- atrophy and absence of CD4, CD8 double positive cells in doxycycline treated mice
- immune system phenotype
- spleen atrophy (MGI Ref ID J:98920)
- in doxycycline treated mice
- thymus atrophy (MGI Ref ID J:98920)
- atrophy and absence of CD4, CD8 double positive cells in doxycycline treated mice
- tumorigenesis
- skin tumor (MGI Ref ID J:98920)
- in doxycycline treated mice, tumors originating from the outer-root-sheath progenitors and invading the subcutaneous layer are seen
Research Applications
This mouse can be used to support research in many areas including:
Cancer Research
Other
Cell Biology Research
Transcriptional Regulation
Research Tools
Cancer Research (Tetop Tet System)
Genetics Research (Mutagenesis and Transgenesis: Tetop Tet System)
Genetics Research (Mutagenesis and Transgenesis: transcriptional activation)
Tet Expression Systems (tTA/rtTA Expressing Strains)
Tet Expression Systems (tTA/rtTA Responsive Strains)
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Col1a1tm2(tetO-Pou5f1)Jae | ||
|---|---|---|---|
| Allele Name | targeted mutation 2, Rudolf Jaenisch | ||
| Common Name(s) | Col1a1::TetOP-Oct4; | ||
| Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 | ||
| ES Cell Line Name | v6.5 | ||
| ES Cell Line Strain | (C57BL/6 x 129S4/SvJae)F1 | ||
| Expressed Gene | Pou5f1, POU domain, class 5, transcription factor 1, mouse, laboratory | ||
| General Note | This mutation is present in the same mouse line carrying the Gt(ROSA)26Sortm1(rtTA*M2)Jae mutation. Expression is induced by doxycycline administration. | ||
| Molecular Note | ES cells containing the Gt(ROSA)26Sortm1(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. [MGI Ref ID J:98920] | ||
| Gene Symbol and Name | Col1a1, collagen, type I, alpha 1 | ||
| Chromosome | 11 | ||
| Gene Common Name(s) | COLIA1; Col1a-1; Cola-1; Cola1; Moloney leukemia virus 13; Mov-13; OI4; | ||
| Allele Symbol | Gt(ROSA)26Sortm1(rtTA*M2)Jae | ||
| Allele Name | targeted mutation 1, Rudolf Jaenisch | ||
| Common Name(s) | Gt(ROSA)26Sortm1(M2rtTA)Jae; R26-M2rtTA; R26-rtTA; | ||
| Mutation Made By | Rudolf Jaenisch, Whitehead Institute (MIT) | ||
| Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 | ||
| ES Cell Line Name | v6.5 | ||
| ES Cell Line Strain | (C57BL/6 x 129S4/SvJae)F1 | ||
| 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. | ||
| Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano | ||
| Chromosome | 6 | ||
| Gene Common Name(s) | AV258896; Gtrgeo26; Gtrosa26; R26; ROSA26; beta geo; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; | ||
| General Note |
This mutation is always used in combination with the Col1a1 | ||
| Molecular Note | An optimized form of reverse tetracycline controlled transactivator (rtTA-M2) was inserted downstream of the Gt(ROSA)26Sor promoter. 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 transactivatory protein has increased doxycycline sensitivity. Mice have widespread expression of the rtTA-M2 protein. Doxycycline treatment induces expression of Pou5f1 from the Col1a1 locus under the control of a tetracycline-responsive element in these mice. [MGI Ref ID J:98920] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Col1a1tm2(tetO-Pou5f1)Jae, SEP PCR, vers. 1
Gt(ROSA)26Sortm1(rtTA*M2)Jae, SEP PCR, vers. 1
Helpful Links
Optimizing PCR Protocols
References
References
Selected Reference(s)
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):465-77. [PubMed: 15882627] [MGI Ref ID J:98920]
Additional References
Gt(ROSA)26Sortm1(rtTA*M2)Jae related
Brennand K; Huangfu D; Melton D. 2007. All beta Cells Contribute Equally to Islet Growth and Maintenance. PLoS Biol 5(7):e163. [PubMed: 17535113] [MGI Ref ID J:124045]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
Pricing for USA, Canada and Mexico shipping destinations
View International Pricing
Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $236.40 Female or Male Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae $291.90 Female or Male Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Homozygous for Col1a1tm2(tetO-Pou5f1)Jae *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $472.80 Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae x Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae
Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement. |
|---|---|
| Supply Notes |
|
Pricing for International shipping destinations
View USA, Canada and Mexico Pricing
Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $307.40 Female or Male Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae $379.50 Female or Male Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Homozygous for Col1a1tm2(tetO-Pou5f1)Jae *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $614.70 Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae x Homozygous for Gt(ROSA)26Sortm1(rtTA*M2)Jae, Heterozygous for Col1a1tm2(tetO-Pou5f1)Jae
Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement. |
|---|---|
| Supply Notes |
|
Control Information
| Control | ||
|---|---|---|
| 101043 B6129SF1/J | (approximate) | |
| 101045 B6129SF2/J | (approximate) | |
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
General Terms and Conditions
View JAX® Mice & Services Conditions of Use.
Use of the Tet-System may require a license, see Licenses for Strains Using TET-System Technology.
For additional Licensing and Use Restrictions view the link(s) below:
- Use of MICE by companies or for-profit entities requires a license prior to shipping.
- Use of MICE by companies or for-profit entities requires a license.
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.Ordering and Purchasing Information
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