This R26STOPrtTA mutant mouse strain combines two popular conditionally-regulated expression methodologies (Tet Expression and Cre Recombinase) providing a tool for doxycycline-inducible spatiotemporal and lineage specific expression of genes.
Frank Costantini, Columbia University Medical Center
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Transactivator) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
These mice contain the rtTAS*M2, or rtTA2S-M2, an optimized form of reverse tetracycline regulated transactivator, gene inserted into the Gt(ROSA)26Sor locus. Expression of the rtTAS*M2 is blocked by a loxP-flanked STOP fragment placed between the rtTAS*M2 sequence and the Gt(ROSA)26Sor promoter. When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will express rtTAS*M2, reverse tetracycline regulated transactivator, in the cre expressing cells. When bred to mice that also carry a gene of interest under the regulatory control of a tetracycline-responsive promoter element (tetO), expression of the gene of interest may be regulated by the tetracycline analog, doxycycline (dox). In the presence of dox, transcription of the target gene is induced in cells where rtTA is expressed. Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.
A targeting vector was used to insert a loxP flanked STOP sequence followed by an optimized reverse tetracycline-controlled transactivator, rtTAS*M2, or rtTA2S-M2, into Gt(ROSA)26Sor.
The construct was electroporated into 129S6/SvEvTac derived CSL3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6J mice for an unknown number of generations. Homozygote males were then crossed to wildtype 129S6/SvEvTac females.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Allele Name | targeted mutation 1, Wei Hsu |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Transactivator) |
Allele Synonym(s) | Gt(ROSA)26Sortm1(xstpx-rtTA2S*M2)Whsu; R26STOPrtTA |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Site of Expression | When mice carrying this allele are bred to mice that express Cre recombinase, the resulting offspring will express rtTAS*M2, reverse tetracycline regulated transactivator, in the cre expressing cells. When bred to mice that also carry a gene of interest under the regulatory control of a tetracycline-responsive promoter element (tetO), expression of the gene of interest may be regulated by the tetracycline analog, doxycycline (dox). In the presence of dox, transcription of the target gene is induced in cells where rtTA is expressed. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | A synthetic modified version of the rtTA gene preceded by a loxP flanked STOP sequence was inserted into the Gt(ROSA)26Sor locus. This version of rtTA contains 5 amino acid changes in the TetR moiety (S12G, E19G, A56P, D148E, and H179R) and a synthetic optimized transactivating domain. This rtTA has reduced basal activity and enhanced doxycycline sensitivity compared to wild-type rtTA. |
Mutations Made By | Frank Costantini, Columbia University Medical Center |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the B6;129S6-Gt(ROSA)26Sortm1(rtTA2S*M2)Whsu/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #016999 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Gt(ROSA)26Sor<tm1(xstpx-rtTA2S*M2)Whsu> |
Frozen Mouse Embryo | B6;129S6-Gt(ROSA)26Sor<tm1(rtTA2S*M2)Whsu>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S6-Gt(ROSA)26Sor<tm1(rtTA2S*M2)Whsu>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S6-Gt(ROSA)26Sor<tm1(rtTA2S*M2)Whsu>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129S6-Gt(ROSA)26Sor<tm1(rtTA2S*M2)Whsu>/J Frozen Embryo | $3373.50 |
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