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 R26M2rtTA targeted mutation (an optimized form of reverse tetracycline controlled transactivator [rtTA-M2] downstream of the Gt(ROSA)26Sor promoter) and the Col1a1TetOP-Cre targeted mutation (a tetracycline responsive element-driven Cre recombinase targeted to the collagen type I alpha 1 locus).
Hanno R Hock, Massachusetts General Hospital
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Transactivator) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Recombinase-expressing, Inducible) | Col1a1 | collagen, type I, alpha 1 |
R26-M2rtTA::TetOP-Cre mice harbor the Rosa26-rtTA targeted mutation and the Col1a1TetOP-Cre targeted mutation. Rosa26-rtTA has an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) downstream of the Gt(ROSA)26Sor promoter. Col1a1TetOP-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.
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.
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 | 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. |
Allele Name | targeted mutation 1, Rudolf Jaenisch |
---|---|
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, Hanno Hock |
---|---|
Allele Type | Targeted (Recombinase-expressing, Inducible) |
Allele Synonym(s) | R26M2rtTA;Col1a1TetOP-Cre; R26-M2rtTA::TetOP-Cre; TetOP-Cre |
Gene Symbol and Name | Col1a1, collagen, type I, alpha 1 |
Gene Synonym(s) | |
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 (Col1a1tm13(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 Col1a1tm13(neo/hygro)Jae ES cells also contain the Gt(ROSA)26Sortm1(rtTA*M2)Jae allele. The twice-targeted ES cells were used to generate chimeras and germline animals. By treating this mouse line with doxycycline, rtTA*M2 binds the TetOP promoter and cre is expressed. |
When maintaining a live colony, mice that are homozygous for both mutations may be bred together.
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.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Heterozygous for Col1a1<tm1(tetO-cre)Haho> |
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 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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.