Overview

Also Known As:Rosa26-rtTA::Col1a1-tetO-H2B-mCherry

These double mutant Rosa26-rtTA::Col1a1-tetO-H2B-mCherry mice harbor the Rosa26-rtTA targeted mutation (an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) downstream of the Gt(ROSA)26Sor promoter) and the Col1a1-tetO-H2B-mcherry targeted mutation (a tet-responsive element (tetO) and a histone H2B-mCherry fusion protein, targeted to the collagen, type I, alpha 1 (Col1a1) locus). These Rosa26-rtTA::Col1a1-tetO-H2B-mCherry mice may be useful for doxycycline-inducible studies which utilize the rtTA/tet-O (tet-on/TRE) system for visualizing pluripotent cells.

Donating Investigator

IMR Colony, The Jackson Laboratory

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^{tm1(tetO-mCherry)Eggn}

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter, Null/Knockout, RMCE-ready)	Col1a1	collagen, type I, alpha 1

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Research Applications

Cancer Research
Cell Biology Research
Research Tools

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The Rosa26-rtTA::Col1a1-tetO-H2B-mCherry double mutant mice were generated by breeding Rosa26-rtTA mice (from Stock No. 006965) with Col1a1-tetO-H2B-mCherry mice (from Stock No. 014592). The Rosa26-rtTA targeted mutation has an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) downstream of the Gt(ROSA)26Sor promoter. The Col1a1-tetO-H2B-mcherry targeted mutation contains a tet-responsive element (tetO) and a histone H2B-mCherry fusion protein, targeted to the collagen, type I, alpha 1 (Col1a1) locus.

Rosa26-rtTA::Col1a1-tetO-H2B-mCherry double mutant mice that are heterozygous for both targeted mutations are viable and fertile (mice homozygous for both targeted mutations were not generated by the donating investigator but are expected to be viable and fertile as well). These double mutant Rosa26-rtTA::Col1a1-tetO-H2B-mCherry mice have widespread expression of the optimized form of reverse tetracycline-controlled transactivator (rtTA-M2) protein directed to multiple tissues by the Gt(ROSA)26Sor promoter. In the absence of the tetracycline analog doxycycline (dox), expression of the dox-inducible histone H2B-mCherry fusion protein cassette from the Col1a1 locus is not detected. Following dox administration, double mutant mice express the H2B-mCherry fusion protein in embryonic stem (ES) cells, and differentiated trophoblast stem (TS) cells. These double mutant Rosa26-rtTA::Col1a1-tetO-H2B-mCherry mice may be useful for doxycycline-inducible studies which utilize the rtTA/tet-O (tet-on/TRE) system for visualizing pluripotent cells. Mice homozygous for the Col1a1-tetO-H2B-mCherry mutation may exhibit a subtle kink in the tail.

Development

These mice harbor the Rosa26-rtTA targeted mutation and the Col1a1-tetO-H2B-mCherry targeted mutation. The Rosa26-rtTA targeted mutation was created by Dr. Rudolf Jaenisch (Whitehead Institute, MIT) and sent to The Jackson Laboratory Repository as Stock No. 006965. The Col1a1-tetO-H2B-mCherry targeted mutation was created by Dr. Kevin Eggan (Harvard University) and sent to The Jackson Laboratory Repository as Stock No. 014592. The Rosa26-rtTA::Col1a1-tetO-H2B-mCherry double mutant mice were generated at The Jackson Laboratory Repository by breeding Rosa26-rtTA mice (from Stock No. 006965) with Col1a1-tetO-H2B-mCherry mice (Stock No. 014592). Double mutant mice were then bred together to maintain the colony (Stock No. 014602).

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the F3-F5 generation living colony at The Jackson Laboratory Repository. This revealed 3 of 32 markers, one each on chromosomes 7, 11 and 13, that were not fixed for C57BL/6 allele-type (e.g., still segregating for 129S4 allele-type markers) in some animals. Collectively, these data suggest the mice are >90% C57BL/6J with some (but not all) animals still segregating for 129S4 regions from the Col1a1-tetO-H2B-mCherry donor strain.

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	RFP, Red Fluorescent Protein, coral
Site of Expression	Following doxycycline administration, H2B-mCherry fusion protein is expressed in embryonic stem (ES) cells, and differentiated trophoblast stem (TS) cells.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Egli D; Rosains J; Birkhoff G; Eggan K. 2007. Developmental reprogramming after chromosome transfer into mitotic mouse zygotes. Nature 447(7145):679-85PubMed: 17554301 MGI: J:167431
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^{tm1(tetO-mCherry)Eggn}

Allele Symbol: Col1a1^{tm1(tetO-mCherry)Eggn}

Allele Name	targeted mutation 1, Kevin Eggan
Allele Type	Targeted (Reporter, Null/Knockout, RMCE-ready)
Allele Synonym(s)	Col1a1TetoH2BCherry
Gene Symbol and Name	Col1a1, collagen, type I, alpha 1
Gene Synonym(s)
Promoter	tetO, tet operator,
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Following doxycycline administration, H2B-mCherry fusion protein is expressed in embryonic stem (ES) cells, and differentiated trophoblast stem (TS) cells.
Strain of Origin	(C57BL/6 x 129S4/SvJae)F1
Chromosome	11
General Note	The allele was made in KH2 cells that carry Gt(ROSA)26Sor^{tm1(rtTAM2)Jae} and Col1a1^{tm13(neo/hygro)Jae}. This allele was generated with Col1a1^{tm13(neo/hygro*)Jae}.
Molecular Note	Using KH2 ES cells inserted a tetO-mCherry "flip-in plasmid" into the 3'UTR. The tetO-mCherry "flip-in plasmid" contained a splice acceptor-double polyA sequence and the tetracycline responsive element (tetO) upstream of a histone H2B minimal promoter-mCherry fluorescent protein fusion protein.
Mutations Made By	Kevin Eggan, Harvard University

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

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

Mammalian Phenotype Terms by Genotype

References

Egli D; Rosains J; Birkhoff G; Eggan K. 2007. Developmental reprogramming after chromosome transfer into mitotic mouse zygotes. Nature 447(7145):679-85PubMed: 17554301 MGI: J:167431
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^{tm1(tetO-mCherry)Eggn} related

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

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Gt(rosa)26sor Probe
- Standard PCR:Gt(ROSA)26Sor
- Separated MCA:Gt(ROSA)26Sor STD
- Standard PCR:Gt(ROSA)26Sor STD
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice that are homozygous for both mutations may be bred together. Mice homozygous for the Col1a1-tetO-H2B-mCherry mutation may exhibit a subtle kink in the tail.
- Additional Breeding and Husbandry Support
Citation
When using the Rosa26-rtTA::Col1a1-tetO-H2B-mCherry mouse strain in a publication, please cite the originating article(s) and include JAX stock #014602 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 for Col1a1<tm1(tetO-mCherry)Eggn>

Related Products and Services

Frozen Mouse Embryo	B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<t Frozen Embryos	$2595.00
Frozen Mouse Embryo	B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<t Frozen Embryos	$2595.00
Frozen Mouse Embryo	B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<t Frozen Embryos	$3373.50
Frozen Mouse Embryo	B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<t Frozen Embryos	$3373.50

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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:Rosa26-rtTA::Col1a1-tetO-H2B-mCherry

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

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

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

Col1a1tm1(tetO-mCherry)Eggn

Allele Symbol: Col1a1tm1(tetO-mCherry)Eggn

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Col1a1tm1(tetO-mCherry)Eggn 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

Related Products and Services

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^{tm1(tetO-mCherry)Eggn}

Allele Symbol: Col1a1^{tm1(tetO-mCherry)Eggn}

Additional - Col1a1^{tm1(tetO-mCherry)Eggn} related

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