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.
IMR Colony, The Jackson Laboratory
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Transactivator) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, Null/Knockout, RMCE-ready) | Col1a1 | collagen, type I, alpha 1 |
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.
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.
Expressed Gene | rtTA, reverse tetracycline-controlled transactivator, E. coli |
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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. |
Allele Name | targeted mutation 1, Rudolf Jaenisch |
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Allele Type | Targeted (Transactivator) |
Allele Synonym(s) | Gt(ROSA)26Sortm1(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 |
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 |
Allele Name | targeted mutation 1, Kevin Eggan |
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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)26Sortm1(rtTA*M2)Jae and Col1a1tm2(tetO-Pou5f1)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 |
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.
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.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm1(rtTA*M2)Jae>, Heterozygous for Col1a1<tm1(tetO-mCherry)Eggn> |
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 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.
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