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| These CAG-stop-tTA2 transgenic mice have a loxP-flanked transcriptional STOP cassette preventing transcription of the downstream modified tetracycline-regulated transactivator (tTA2). Applying both Cre-lox and Tet-Off technologies, these CAG-stop-tTA2 transgenic mice may be useful to generate compound mutant mice in which expression of a tetracycline-responsive promoter element (TRE or tetO)-driven gene of interest can be both directed to the cell types defined by the chosen Cre recombinase expression, as well as turned off by the addition of tetracycline (or its analog doxycycline [dox]). | |||||||||||||||||||
- Description
- Disease & phenotype
- Genes & alleles
- Genotyping
- Health & care
- References
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Description
The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.
Strain Information
Former Names STOCK Tg(ACTB-tTA2,-lacZ)1Luo/J (Changed: 13-JAN-11 ) Type Mutant Stock; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Species laboratory mouse Donating Investigator Dr. Liqun Luo, Stanford University, HHMI Description
Homozygous CAG-stop-tTA2 transgenic mice are viable and fertile. CAG-stop-tTA2 transgenic mice harbor the ii-CAG-stop-tTA2-IRES-tau-lacZ-ii transgene; designed with a loxP-flanked transcriptional STOP cassette preventing transcription of the downstream modified tetracycline-regulated transactivator (tTA2). The ii-CAG-stop-tTA2-IRES-tau-lacZ-ii transgene is flanked by two copies of the chicken β-globin HS4 insulator on each side to preserve expression fidelity (see additional information below). When bred to mice that express a tamoxifen-inducible Cre recombinase (CreERT2), administration of tamoxifen to the double mutant offspring allows the CreERT2 fusion protein to enter the nucleus of the cre-expressing cells; this deletes the STOP cassette and results in expression of tTA2. The donating investigator reports that tau-lacZ fusion protein expression in the tamoxifen-treated double mutant offspring is faint. Of note, the donating investigator has not confirmed if breeding CAG-stop-tTA2 transgenic mice with mice expressing constitutively active Cre recombinase results in tTA2 expression in the double mutant offspring.Applying both Cre-lox and Tet-Off technologies, these CAG-stop-tTA2 transgenic mice may be useful to generate compound mutant mice in which expression of a tetracycline-responsive promoter element (TRE or tetO)-driven gene of interest can be both directed to the cell types defined by the chosen Cre recombinase expression, as well as turned off by the addition of tetracycline (or its analog doxycycline [dox]).
These CAG-stop-tTA2 transgenic mice may be used to design a tripartite system to express fluorescently tagged synaptic proteins with both spatial and temporal control. When CAG-stop-tTA2 transgenic mice are bred with TRE-Bi-SG-T transgenic mice (Stock No. 012345), the resulting CAG-stop-tTA2::TRE-Bi-SG-T double mutant mice allow tdTomato and synaptophysin/GFP fusion protein expression to be determined by the tissue-specific tamoxifen-inducible Cre recombinase expression of a third strain of the researchers choosing. If such CAG-stop-tTA2::TRE-Bi-SG-T double mutant mice were bred with CaMKII-CreER mice (Stock No. 012362), the resulting tamoxifen-treated triple mutant mice should exhibit red-labeling of whole cortical/hippocampal/striatal neurons and green-labeling of their presynaptic terminals in the absence of dox. Similarly, if CAG-stop-tTA2::TRE-Bi-SG-T double mutant mice were bred with GAD2-CreER mice (Stock No. 010702), the resulting tamoxifen-treated triple mutant mice should exhibit red-labeling of whole GABAergic interneurons and green-labeling of their presynaptic terminals in the absence of dox.
The CAG-stop-tTA2 transgene is flanked by chicken β-globin HS4 insulators. The insulators are reported to preserve expression fidelity from transgene integration/position effects by blocking expansion of heterochromatin into the transgene (potentially preventing it from being silenced), as well as decrease the basal expression/increase the inducibility of TREs.
Development
The ii-CAG-stop-tTA2-IRES-tau-lacZ-ii transgene was designed with the human cytomegalovirus (CMV) enhancer/chicken beta-actin core promoter (pCA), a loxP-flanked neomycin resistance gene (neor) with transcriptional STOP cassette, a modified tetracycline-regulated transactivator gene (tTA2), an internal ribosome entry site (IRES), and a microtubule-associated protein tau with β-galactosidase fusion protein (tau-lacZ). The transgene was further modified to have two copies of a ~250 bp chicken β-globin HS4 insulator sequence (separated by an ApaLI restriction site) on each side of the transgene. The 5' insulator pair is flanked by Saw1 and Kpn1 restriction sites, while the 3' insulator pair is flanked by Spe1 and Asc1 restriction sites. The resulting ii-CAG-stop-tTA2-IRES-tau-lacZ-ii transgene was microinjected into the pronuclei of zygotes from DBA2 oocytes fertilized with C57BL6 sperm. Founder mice determined to harbor the neor gene (via PCR genotyping) were bred to β-actin-CreER mutant mice on a mixed genetic background (CD1, 129, C57BL/6) to test them for expression and to establish founder lines. Mice from founder line 1 were found to have 2-3 copies of the transgene. The CAG-stop-tTA2 transgenic mice were bred to other mutant/transgenic mice on mixed genetic backgrounds (mixed CD1, 129, C57BL/6 and/or FVB) for several generations. CAG-stop-tTA2 transgenic mice were also bred together and to outbred CD1 for at least five generations prior to sending to The Jackson Laboratory Repository. Upon arrival, CAG-stop-tTA2 transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| Considerations for Choosing Controls | ||
Related Strains
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Additional Web Information
Introduction to Cre-lox technology
Tet Expression Systems
Visit the Alzheimer's Disease Mouse Model Resource site for helpful information on Alzheimer's Disease and research resources.
Visit the Parkinson's Disease Resource site for helpful information on Parkinson's and research resources.
Phenotype
Phenotype Information
- Potential model based on transgenic expression of an ortholog of a human gene that is associated with this disease. Phenotypic similarity to the human disease has not been tested. Frontotemporal Dementia; FTD (MAPT)
Parkinson Disease, Late-Onset; PD (MAPT)
Parkinson-Dementia Syndrome (MAPT)
Pick Disease of Brain (MAPT)
Supranuclear Palsy, Progressive, 1; PSNP1 (MAPT)
This mouse can be used to support research in many areas including:
MAPT relatedNeurobiology Research
Cre-lox System
loxP-flanked Sequences
Tet Expression System
tTA/rtTA Expressing Strains
Research Tools
Cancer Research
Tetop Tet System
Cardiovascular Research
Cre-lox System
Tetop Tet System
Cre-lox System
loxP-flanked Sequences
Developmental Biology Research
Cre-lox System
Diabetes and Obesity Research
loxP
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Tetop Tet System
Tissue/Cell Markers
Tissue/Cell Markers: Cre-lox System
Neurobiology Research
Tetop Tet System
Reproductive Biology Research
Cre-lox System
Tetop Tet System
Tet Expression Systems
tTA/rtTA Expressing Strains
Neurobiology Research
Alzheimer's Disease
Parkinson's Disease
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Tg(ACTB-tTA2,-MAPT/lacZ)1Luo | ||
|---|---|---|---|
| Allele Name | transgene insertion 1, Liqun Luo | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | CAG-stop-tTA2; CAG-stop-tTA2-IRES-tau-lacZ; Tg(ACTB-tTA2,-lacZ)1Luo; ii-CAG-stop-tTA2-IRES-tau-lacZ-ii; | ||
| Mutation Made By | Dr. Liqun Luo, Stanford University, HHMI | ||
| Strain of Origin | DBA/2 | ||
| Site of Expression | these CAG-stop-tTA2 transgenic mice may be useful to generate compound mutant mice in which expression of a tetracycline-responsive promoter element (TRE or tetO)-driven gene of interest can be both directed to the cell types defined by the chosen Cre recombinase expression, as well as turned off by the addition of tetracycline (or its analog doxycycline [dox]). | ||
| Expressed Gene | lacZ, beta-galactosidase, E. coli | ||
| Expressed Gene | MAPT, microtubule-associated protein tau, human | ||
| Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli | ||
| The tetracycline-resistance gene (TetR), arose from chemically mutated Escherichia coli genome which was screened for tetracycline dependence (Gossen and Bujard, 1992). TetR was fused at the C-terminus with the viral co-activator, virion protein 16 of the herpes simplex virus (VP-16). The tetracycline-inhibitable transcription factor is a component of a bigenic system that allows doxycycline (a tetracycline analog) regulatable expression of genes that are under the direction of the tetracycline responsive promoter (TetOp)promoter. | |||
| Promoter | ACTB, actin, beta, chicken | ||
| Promoter | CMV, cytomegalovirus, human | ||
| Molecular Note | The ii-CAG-stop-tTA2-IRES-tau-lacZ-ii transgene was designed with the human cytomegalovirus (CMV) enhancer/chicken beta-actin core promoter (pCA), a loxP-flanked neomycin resistance gene (neor) with transcriptional STOP cassette, a modified tetracycline-regulated transactivator gene (tTA2), an internal ribosome entry site (IRES), and a bovine microtubule-associated protein tau with beta-galactosidase fusion protein (tau-lacZ). The transgene was further modified to have two copies of a ~250 bp chicken beta-globin HS4 insulator sequence (separated by an ApaLI restriction site) on each side of the transgene. The 5' insulator pair is flanked by Saw1 and Kpn1 restriction sites, while the 3' insulator pair is flanked by Spe1 and Asc1 restriction sites. [MGI Ref ID J:167275] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(ACTB-tTA2,-lacZ), Separated PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
Miyamichi K; Amat F; Moussavi F; Wang C; Wickersham I; Wall NR; Taniguchi H; Tasic B; Huang ZJ; He Z; Callaway EM; Horowitz MA; Luo L. 2010. Cortical representations of olfactory input by trans-synaptic tracing. Nature :. [PubMed: 21179085] [MGI Ref ID J:167275]
Health & husbandry
The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.
Health & Colony Maintenance Information
Animal Health Reports
Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200.Colony Maintenance
Breeding & Husbandry When maintaining a live colony, hemizygous mice may be bred together, to wildtype (noncarrier) mice from the colony, or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator reports that homozygous mice are viable and fertile, and breeds homozygous mice together.
Pricing and Purchasing
Pricing, Supply Level & Notes, Controls
| Pricing for USA, Canada and Mexico shipping destinations |
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Cryopreserved
Cryopreserved Mice - Ready for Recovery
Animals Provided
Price (US dollars $) Cryorecovery* $1980.00 At least two mice that carry the mutation (if it is a mutant strain) will be provided. Their genotypes may not reflect those discussed in the strain description. Please inquire for possible genotypes and see additional details below.
Standard Supply
Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.
Supply Notes
- Cryorecovery - Standard.
Progeny testing is not required.
The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided, their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders are needed. Animals typically ship between 11 and 14 weeks from the date of your order. If a second cryorecovery is needed in order to provide the minimum number of animals, animals will ship within 25 weeks. IMPORTANT NOTE: The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory 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. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 (from U.S.A., Canada or Puerto Rico only) or 1-207-288-5845 (from any location).
| Pricing for International shipping destinations |
|
Cryopreserved
Cryopreserved Mice - Ready for Recovery
Animals Provided
Price (US dollars $) Cryorecovery* $2574.00 At least two mice that carry the mutation (if it is a mutant strain) will be provided. Their genotypes may not reflect those discussed in the strain description. Please inquire for possible genotypes and see additional details below.
Standard Supply
Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.
Supply Notes
- Cryorecovery - Standard.
Progeny testing is not required.
The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided, their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders are needed. Animals typically ship between 11 and 14 weeks from the date of your order. If a second cryorecovery is needed in order to provide the minimum number of animals, animals will ship within 25 weeks. IMPORTANT NOTE: The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory 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. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 (from U.S.A., Canada or Puerto Rico only) or 1-207-288-5845 (from any location).
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Standard Supply
Cryopreserved. Ready for recovery. Please refer to pricing and supply notes on the strain data sheet for further information.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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