Overview

Also Known As: R26-M2rtTA::TetOP-Cre

These double mutant R26-M2rtTA::TetOP-Cre mice utilize the rtTA/tetO (tet-on/TRE) system to allow highly-efficient, doxycycline-inducible Cre recombinase expression in hematopoietic stem cells. These mice harbor the R26^M2rtTA targeted mutation (an optimized form of reverse tetracycline controlled transactivator [rtTA-M2] downstream of the Gt(ROSA)26Sor promoter) and the Col1a1^TetOP-Cre targeted mutation (a tetracycline responsive element-driven Cre recombinase targeted to the collagen type I alpha 1 locus).

Donating Investigator

Hanno R Hock, Massachusetts General Hospital

Genetic overview

Genetic Background	Generation

Col1a1^{tm1(tetO-cre)Haho}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing, Inducible)	Col1a1	collagen, type I, alpha 1

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

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

View Genetics

Research Applications

Research Tools
Cell Biology Research
Cancer Research
Neurobiology Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

R26-M2rtTA::TetOP-Cre mice harbor the Rosa26-rtTA targeted mutation and the Col1a1^TetOP-Cre targeted mutation. Rosa26-rtTA has an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) downstream of the Gt(ROSA)26Sor promoter. Col1a1^TetOP-Cre contains a tetracycline responsive element-driven Cre recombinase targeted to the collagen type I alpha 1 locus.

Mice homozygous for both targeted mutations (homozygous R26-M2rtTA::TetOP-Cre mice) or heterozygous for both targeted mutations (heterozygous R26-M2rtTA::TetOP-Cre mice) are viable and fertile. Following administration of the tetracycline analog doxycycline (dox), the donating investigator reports sufficient Cre recombinase activity is observed for both homozygous R26-M2rtTA::TetOP-Cre mice and heterozygous R26-M2rtTA::TetOP-Cre mice. Specifically, homozygous R26-M2rtTA::TetOP-Cre mice exhibit Cre recombinase activity in hematopoietic stem cells at high efficiency (close to 100% for floxed alleles that face no selection in hematopoietic stem cells upon recombination); and dox-inducible Cre recombinase activity in adult tissues is reported at ~30-50% efficiency. In the absence of dox, heterozygous R26-M2rtTA::TetOP-Cre mice exhibit no Cre recombinase expression from the Col1a1 locus. Homozygous R26-M2rtTA::TetOP-Cre mice have a low degree of Cre recombinase expression prior to dox administration (causing low levels of recombination with some floxed alleles; less than 10% germline excision occurs). In vitro, a minor reduction in the rate of hematopoietic progenitor colony growth is observed for homozygous R26-M2rtTA::TetOP-Cre mice, but not heterozygous R26-M2rtTA::TetOP-Cre mice.

Development

The R26-M2rtTA targeted mutation was created by Dr. Rudolf Jaenisch (Whitehead Institute, MIT) with 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 endogenous promoter at the Gt(ROSA)26Sor locus. The targeting vector was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Next, these targeted ES cells (termed V19 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. Next, Dr. Hanno R. Hock (Massachusetts General Hospital) replaced the frt-flanked PGK-Neo in the 3' UTR of the Col1a1 locus with a TetOP-Cre sequence. This was performed by injecting a "TetOP-Cre flip-in plasmid" (containing a splice acceptor-polyA sequence and the tetracycline operator elements fused to a CMV minimal promoter [TRE or tetO] upstream of a Cre recombinase cassette [from pTurbo-cre; Genbank AF334827.1]), along with a Flpe-expressing plasmid (to facilitate TetOP-Cre 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 TetOP-Cre in the Col1a1 locus (R26-M2rtTA::TetOP-Cre) were injected into B6D2F1 tetraploid blastocysts. The double mutant chimeric males were bred with C57BL/6J females to generate the colony. Double mutant pups on the resulting C57BL/6;129 mixed genetic background were bred together for several generations, and then sent to The Jackson Laboratory Repository in 2014. Upon arrival, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation.

Expression Data

Expressed Gene	cre, cre recombinase, bacteriophage P1
Site of Expression	When combined with a reverse tetracycline controlled transactivator, cre recombinase expression can be induced in hematopoietic stem cells by doxycycline administration.
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.

Control Suggestions

Depending upon the experiment, the following mouse lines may also be appropriate controls:
C57BL/6-congenic R26-M2rtTA (Stock No. 006965)
R26-M2rtTA::TetOP-H2B-GFP (Stock No. 016836)

Approximate Controls

101045 B6129SF2/J

Additional Information

Considerations for Choosing Controls

Selected References

Beard C; Hochedlinger K; Plath K; Wutz A; Jaenisch R. 2006. Efficient method to generate single-copy transgenic mice by site-specific integration in embryonic stem cells. Genesis 44(1):23-8PubMed: 16400644 MGI: J:159351
Foudi A; Hochedlinger K; Van Buren D; Schindler JW; Jaenisch R; Carey V; Hock H. 2009. Analysis of histone 2B-GFP retention reveals slowly cycling hematopoietic stem cells. Nat Biotechnol 27(1):84-90PubMed: 19060879 MGI: J:172421
Foudi A; Kramer DJ; Qin J; Ye D; Behlich AS; Mordecai S; Preffer FI; Amzallag A; Ramaswamy S; Hochedlinger K; Orkin SH; Hock H. 2014. Distinct, strict requirements for Gfi-1b in adult bone marrow red cell and platelet generation. J Exp Med 211(5):909-27PubMed: 24711581 MGI: J:207446
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

View All References

Genetics

Col1a1^{tm1(tetO-cre)Haho}

Allele Symbol: Col1a1^{tm1(tetO-cre)Haho}

Allele Name	targeted mutation 1, Hanno Hock
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	Col1a1^{tm1(tetO-cre)Haho}; targeted mutation 1, Hanno Hock
Gene Symbol and Name	Col1a1, collagen, type I, alpha 1
Gene Synonym(s)	Cola1; Cola1; COLIA1; Mov-13; Col1a-1; Cola-1; Mov-13; Moloney leukemia virus 13; OI4; Col1a-1; Cola-1; OI3; EDSC; OI1; OI2; EDSARTH1
Promoter	tetO, tet operator,
Expressed Gene	cre, cre recombinase, bacteriophage P1
Site of Expression	When combined with a reverse tetracycline controlled transactivator, cre recombinase expression can be induced in hematopoietic stem cells by doxycycline administration.
Strain of Origin	(C57BL/6 x 129S4/SvJae)F1
Chromosome	11
Molecular Note	A "TetOP-Cre flip-in plasmid" (containing a splice acceptor-polyA sequence and the tetracycline operator elements (tet)) fused to a CMV minimal promoter upstream of a cre recombinase cassette) was used to treat ES cells containing an FRT-PGK-neo-hygro-FRT cassette inserted into the 3' UTR of the Col1a1 locus (Col1a1^{tm13(neo/hygro)Jae}). The "flip-in" vector was coinjected with a Flpe-encoding plasmid so that flippase-mediated recombination inserted the tetO-cre sequence into the Col1a1 3' UTR. The Col1a1^{tm13(neo/hygro)Jae} ES cells also contain the Gt(ROSA)26Sor^{tm1(rtTAM2)Jae} allele. The twice-targeted ES cells were used to generate chimeras and germline animals. By treating this mouse line with doxycycline, rtTAM2 binds the TetOP promoter and cre is expressed.

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(rtTA*M2)Jae}; targeted mutation 1, Rudolf Jaenisch
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)	beta geo; ROSA26; R26; Gtrgeo26; Gtrosa26; Gtrgeo26; Gtrosa26; AV258896; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; SETD5-AS1; Thumpd3as1
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	Dr. Rudolf Jaenisch, Whitehead Institute (MIT)

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Research Tools
- Reproductive Biology Research
  - Cre-lox System
- Genetics Research
  - Mutagenesis and Transgenesis: transcriptional activation
  - Mutagenesis and Transgenesis: Tetop Tet System
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers: Cre-lox System
- Neurobiology Research
  - Tetop Tet System
- Tet Expression Systems
  - tTA/rtTA Expressing Strains
  - tTA/rtTA Responsive Strains
- Cre-lox System
  - Cre Recombinase Expression: Inducible
- Developmental Biology Research
  - Cre-lox System
- Cancer Research
  - Tetop Tet System
- Cardiovascular Research
  - Tetop Tet System
Cell Biology Research
- Transcriptional Regulation
Cancer Research
- Other
Neurobiology Research
- Tet Expression System
  - tTA/rtTA Expressing Strains
  - tTA/rtTA Responsive Strains
- Cre-lox System
  - Cre Recombinase expression in neural tissue

Mammalian Phenotype Terms by Genotype

References

Beard C; Hochedlinger K; Plath K; Wutz A; Jaenisch R. 2006. Efficient method to generate single-copy transgenic mice by site-specific integration in embryonic stem cells. Genesis 44(1):23-8PubMed: 16400644 MGI: J:159351
Foudi A; Hochedlinger K; Van Buren D; Schindler JW; Jaenisch R; Carey V; Hock H. 2009. Analysis of histone 2B-GFP retention reveals slowly cycling hematopoietic stem cells. Nat Biotechnol 27(1):84-90PubMed: 19060879 MGI: J:172421
Foudi A; Kramer DJ; Qin J; Ye D; Behlich AS; Mordecai S; Preffer FI; Amzallag A; Ramaswamy S; Hochedlinger K; Orkin SH; Hock H. 2014. Distinct, strict requirements for Gfi-1b in adult bone marrow red cell and platelet generation. J Exp Med 211(5):909-27PubMed: 24711581 MGI: J:207446
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-cre)Haho} related

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

Technical Support

Contact Technical Support

Genotyping Protocols
- Probe: Gt(rosa)26sor^Tm1Sor Probe
- Standard PCR: Col1a1 3'UTR assay2 (flip-in)
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, mice that are homozygous for both mutations may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the R26-M2rtTA::TetOP-Cre mouse strain in a publication, please cite the originating article(s) and include JAX stock #021025 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

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International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service	Genotype	Price
	Heterozygous for Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Heterozygous for Col1a1<tm1(tetO-cre)Haho>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We 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.

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Also Known As: R26-M2rtTA::TetOP-Cre

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Col1a1tm1(tetO-cre)Haho

Allele Symbol: Col1a1tm1(tetO-cre)Haho

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

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

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Col1a1tm1(tetO-cre)Haho 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

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Col1a1^{tm1(tetO-cre)Haho}

Allele Symbol: Col1a1^{tm1(tetO-cre)Haho}

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

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

Additional - Col1a1^{tm1(tetO-cre)Haho} related

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