JAX Mouse Strain Datasheet - 014092

STOCK Tg(ACTB-tTA2,-MAPT/lacZ)1Luo/J

Stock No:: 014092

Transgenic

Cryo Recovery

Overview

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]).

Donating Investigator

Liqun Luo, Stanford University, HHMI

Genetic overview

Genetic Background	Generation

Tg(ACTB-tTA2,-MAPT/lacZ)1Luo

Allele Type
Transgenic (Reporter, Transactivator)

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Research Applications

Neurobiology Research
Research Tools

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Base Price

Starting at:

$2,595.00 Domestic price Cryo Recovery

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Details

Detailed 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 (neo^r) 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 neo^r 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.

Expression Data

Expressed Gene	MAPT, microtubule associated protein tau, human
Expressed Gene	lacZ, beta-galactosidase, E. coli
Expressed Gene	tTA, tetracycline-controlled transactivator, E. coli
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]).

Control Suggestions

Noncarrier

Additional Information

Considerations for Choosing Controls

Selected References

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: J:167275

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Genetics

Tg(ACTB-tTA2,-MAPT/lacZ)1Luo

Allele Symbol: Tg(ACTB-tTA2,-MAPT/lacZ)1Luo

Allele Name	transgene insertion 1, Liqun Luo
Allele Type	Transgenic (Reporter, Transactivator)
Allele Synonym(s)	CAG-stop-tTA2; CAG-stop-tTA2-IRES-tau-lacZ; Tg(ACTB-tTA2,-lacZ)1Luo; ii-CAG-stop-tTA2-IRES-tau-lacZ-ii
Gene Symbol and Name	Tg(ACTB-tTA2,-MAPT/lacZ)1Luo, transgene insertion 1, Liqun Luo
Gene Synonym(s)	Tg(CAG-tTA2,-MAPT/lacZ)1Luo; Tg(CAG-tTA2,-MAPT/lacZ)1Luo
Promoter	ACTB, actin, beta, chicken
Promoter	CMV, cytomegalovirus, human
Expressed Gene	MAPT, microtubule associated protein tau, human
Expressed Gene	lacZ, beta-galactosidase, E. coli
Expressed Gene	tTA, tetracycline-controlled transactivator, E. coli
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]).
Strain of Origin	DBA/2
Chromosome	UN
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.
Mutations Made By	Liqun Luo, Stanford University, HHMI

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Neurobiology 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

Genotype: MAPT related

Neurobiology Research
- Alzheimer's Disease
- Parkinson's Disease

References

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: J:167275

Additional - Tg(ACTB-tTA2,-MAPT/lacZ)1Luo related

Adelson JD; Sapp RW; Brott BK; Lee H; Miyamichi K; Luo L; Cheng S; Djurisic M; Shatz CJ. 2016. Developmental Sculpting of Intracortical Circuits by MHC Class I H2-Db and H2-Kb. Cereb Cortex 26(4):1453-63. PubMed: 25316337 MGI: J:240988

Pricing & Availability

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Cryorecovery - Pricing

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	Please inquire about possible genotypes.

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 10 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.

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.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Tg(ACTB-tTA2,-MAPT/lacZ)1Luo

Allele Symbol: Tg(ACTB-tTA2,-MAPT/lacZ)1Luo

Disease Terms

Research Areas By Genotype

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

Genotype: MAPT related

References

Additional - Tg(ACTB-tTA2,-MAPT/lacZ)1Luo related

Genotyping Protocols

Breeding Considerations

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Progeny testing is not required

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