Overview

Also Known As: Ai62(TITL-tdT)-D or Ai62D

These Ai62(TITL-tdT)-D mice (also called Ai62D) are a fluorescent Cre/Tet reporter line, created by targeted insertion at the Igs7 locus (TIGRE; an intergenic region on mouse chromosome 9 that allows reporter expression to be tightly regulated). After removal of the floxed-STOP cassette by Cre recombinase, they may be used to generate Tet-Off/Tet-On mutant animals with conditional (inducible/reversible) expression of tdTomato.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation

Igs7^{tm62.1(tetO-tdTomato)Hze}

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), Reporter, RMCE-ready)	Igs7	intergenic site 7

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Research Applications

Neurobiology Research
Research Tools

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

These Ai62(TITL-tdT)-D mice (also called Ai62(TITL-tdT)Δhygro or Ai62D) are a fluorescent Cre/Tet reporter line, created by targeted insertion at the Igs7 locus (TIGRE; an intergenic region on mouse chromosome 9 that allows reporter expression to be tightly regulated). Ai62D mice harbor the TIGRE-Ins-TRE-LSL-tdTomato conditional allele designed with a modified Tet response element (TRE or tetO) and loxP-flanked STOP cassette upstream of the tdTomato fluorescent protein. When bred with other mice expressing Cre recombinase, tetracycline-controlled transactivator protein (tTA) and/or reverse tetracycline-controlled transactivator protein (rtTA), tdTomato expression in cells/tissues where the expression patterns of the individual promoters driving Cre recombinase and tTA/rtTA overlap can be regulated with tetracycline or its analog doxycycline (dox). Heterozygous mice are viable and fertile with no reported gross physical or behavioral abnormalities. The donating investigator has not attempted to generate homozygous mice to date (June 2013).

Specifically, the donating investigator reports for triple transgenic mice (Cre+/tTA+/tdTomato+) in the cells expressing both Cre and tTA, tdTomato expression may be detected by direct/native fluorescence and by in situ hybridization (using a tdTomato probe sequence). The donating investigator reports Ai62(TITL-tdT)-D mice have no reported levels of tdTomato expression in absence of tTA. Cre-negative double transgenic mice (Cre-/tTA+/tdTomato+) have no tdTomato expression; although one experimental combination of Cre-/tTA+/tdTomato+ mice were reported to have a few very weakly tdTomato-positive cells in cortex and hippocampus.

For characterization information, see images at the Allen Institute for Brain Science website (Ai62(TITL-tdT) images).

Development

These Ai62(TITL-tdT)-D allele (also called Ai62(TITL-tdT)Δhygro or Ai62D) has a Tet response element, floxed-STOP cassette and tdTomato fluorescent protein, all inserted into the T1 TIGRE locus (an intergenic region on chromosome 9 [between AB124611 and Carm1] determined to have low basal transcriptional activity and allow high Cre and/or Tet inducibility).

Specifically, the pTRE-LSL-tdTomato replacement vector was designed with (from 5' to 3')
an FRT3 site,
two tandem copies of the chicken beta-globin HS4 insulator element (to reduce expression of the reporter gene in the absence of transactivator protein),
a modified Tet response element (TRE or (tetO); described in detail below),
a loxP-flanked STOP cassette (with stop codons in all three reading frames linked to a synthetic pA-hGHpA-PGKpA unit),
a sequence encoding the tdTomato fluorescent protein (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)),
a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability),
a bovine growth hormone polyA signal,
two tandem copies of the chicken beta-globin HS4 insulator element,
an AttB site, a PGK-5’hygro cassette, an RNA splice donor and a FRT5 site.

(129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem cells previously targeted with FRT3::AttB::PGK-neoR-polyA::FRT5::splice acceptor::3’hygro cassette::SV40 polyA:AttP in TIGRE, were re-transfected with the pTRE-LSL-tdTomato replacement vector and Flp recombinase vector for recombinase-mediated cassette exchange (RCME).

Chimeric mice were bred with C57BL/6J-congenic PhiC31 mice (see Stock No. 007743) to remove the AttB/AttP-flanked PGK-hygromycin-SV40polyA cassette and replace it with the recombined AttB/AttP site (AttL).

The Ai62D allele is therefore TIGRE-frt3-Ins-TRE-LSL-tdTomato-WPRE-bGHpA-Ins-AttL.

The resulting Ai62D mice were bred with C57BL/6J wildtype mice for at least three additional generations prior to sending to The Jackson Laboratory Repository in 2013. Upon arrival, mice were bred to C57BL/6J inbred animals (Stock No. 000664) at least once to remove the PhiC31 gene. Both the living Ai62D mouse colony and the cryopreserved sperm are the equivalent of five or more generations backcrossed onto C57BL/6J.

The TRE promoter used here is Tet-responsive P_tight; containing a modified Tet response element (TRE_mod) composed of seven direct 36 bp repeats (each containing the 19 bp tet operator sequence [tetO]). TRE_mod is just upstream of a minimal human cytomegalovirus promoter (P_minCMVΔ), which lacks the enhancer that is part of the complete CMV promoter. Consequently, P_tight is silent in the absence of TetR or rTetR binding to tetO.

Expression Data

Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Expression of tdTomato is under the control of tetO in cells/tissues where promoters driving Cre recombinase are expressed.

Control Suggestions

Wild-type from the colony
000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Madisen L; Garner AR; Shimaoka D; Chuong AS; Klapoetke NC; Li L; van der Bourg A; Niino Y; Egolf L; Monetti C; Gu H; Mills M; Cheng A; Tasic B; Nguyen TN; Sunkin SM; Benucci A; Nagy A; Miyawaki A; Helmchen F; Empson RM; Knopfel T; Boyden ES; Reid RC; Carandini M; Zeng H. 2015. Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron 85(5):942-58PubMed: 25741722 MGI: J:219930

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Genetics

Igs7^{tm62.1(tetO-tdTomato)Hze}

Allele Symbol: Igs7^{tm62.1(tetO-tdTomato)Hze}

Allele Name	targeted mutation 62.1, Hongkui Zeng
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter, RMCE-ready)
Allele Synonym(s)	targeted mutation 62.1, Hongkui Zeng; Igs7^{tm62.1(tetO-tdTomato)Hze}
Gene Symbol and Name	Igs7, intergenic site 7
Gene Synonym(s)	transgene insertion 1, George A Gaitanaris; T1; Tg(tetO-Apoe)1Gaga; Iis7; TIGRE; TRE-Apoe; Tg(tetO-Apoe)1Gaga
Promoter	tetO, tet operator,
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Expression of tdTomato is under the control of tetO in cells/tissues where promoters driving Cre recombinase are expressed.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	9
Molecular Note	The targeting constructed introduced by recombinase mediated cassette exchange at the existing site inserted anan FRT3 site, two tandem copies of the chicken beta-globin HS4 insulator element (to reduce expression of the reporter gene in the absence of transactivator protein), a modified Tet response element (TRE or (tetO); described in detail below), a loxP-flanked STOP cassette (with stop codons in all three reading frames linked to a synthetic pA-hGHpA-PGKpA unit), a sequence encoding the tdTomato fluorescent protein (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, two tandem copies of the chicken beta-globin HS4 insulator element, an AttB site, a PGK-5'hygro cassette, an RNA splice donor and a FRT5 site. PhiC31-mediated recombination removed the hygromycin cassette and replaced it with the recombined AttB/AttP site (AttL). The Ai62D allele is therefore TIGRE-frt3-Ins-TRE-LSL-tdTomato-WPRE-bGHpA-Ins-AttL. The modified TRE promoter used in these transgenic mice is the Tet-responsive P^tight promoter. This P^tight promoter contains a modified Tet response element (TRE^mod), which consists of seven direct repeats of a 36-bp sequence each containing the 19-bp tet operator sequence (tetO). The TRE^mod is just upstream of a minimal human cytomegalovirus (CMV) promoter (P^minCMVDelta), which lacks the enhancer that is part of the complete CMV promoter. Consequently, P>tight< is silent in the absence of binding of TetR or rTetR to the tetO sequences.

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
  - loxP-flanked Sequences: Test/Reporter
- Fluorescent protein expression in neural tissue
- Tet Expression System
  - tTA/rtTA Responsive Strains
Research Tools
- Neurobiology Research
  - Tetop Tet System
- Cre-lox System
  - loxP-flanked Sequences: Test/Reporter
  - loxP-flanked Sequences
- Genetics Research
  - Tissue/Cell Markers: cell marker for bone marrow transplantation
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Mutagenesis and Transgenesis
  - Tissue/Cell Markers: Cre-lox System
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers
  - Mutagenesis and Transgenesis: Tetop Tet System
- Diabetes and Obesity Research
  - loxP
- Developmental Biology Research
  - Cre-lox System
  - transplantation marker for embryonic and adult tissue
- Reproductive Biology Research
  - Cre-lox System
  - transplantation marker for embryonic and adult tissue
  - Tetop Tet System
- Tet Expression Systems
  - tTA/rtTA Responsive Strains
- Fluorescent Proteins
- Cancer Research
  - Tetop Tet System
- Cardiovascular Research
  - Tetop Tet System

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Igs7^{tm62.1(tetO-tdTomato)Hze}/Igs7⁺
involves: 129S6/SvEvTac * C57BL/6NCr

normal phenotype

no abnormal phenotype detected
- mice are viable and fertile with no reported gross physical or behavioral abnormalities

References

Madisen L; Garner AR; Shimaoka D; Chuong AS; Klapoetke NC; Li L; van der Bourg A; Niino Y; Egolf L; Monetti C; Gu H; Mills M; Cheng A; Tasic B; Nguyen TN; Sunkin SM; Benucci A; Nagy A; Miyawaki A; Helmchen F; Empson RM; Knopfel T; Boyden ES; Reid RC; Carandini M; Zeng H. 2015. Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron 85(5):942-58PubMed: 25741722 MGI: J:219930

Additional - Igs7^{tm62.1(tetO-tdTomato)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Standard PCR:Iis7^{tm62.1(tetO-tdTomato)Hze}
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator has not attempted to generate homozygous mice to date (June 2013).
- Additional Breeding and Husbandry Support
Citation
When using the Ai62(TITL-tdT)-D or Ai62D mouse strain in a publication, please cite the originating article(s) and include JAX stock #022731 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

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

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Not-For-Profit & Academic Pricing

Service	Genotype	Price
	Heterozygous or wildtype for Igs7<tm62.1(tetO-tdTomato)Hze>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We 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.

Related Products and Services

Frozen Mouse Embryo	B6.Cg-Igs7<tm62.1(tetO-tdTomato)Hze>/J	$2595.00
Frozen Mouse Embryo	B6.Cg-Igs7<tm62.1(tetO-tdTomato)Hze>/J	$2595.00
Frozen Mouse Embryo	B6.Cg-Igs7<tm62.1(tetO-tdTomato)Hze>/J	$3373.50
Frozen Mouse Embryo	B6.Cg-Igs7<tm62.1(tetO-tdTomato)Hze>/J	$3373.50

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Also Known As: Ai62(TITL-tdT)-D or Ai62D

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Igs7tm62.1(tetO-tdTomato)Hze

Allele Symbol: Igs7tm62.1(tetO-tdTomato)Hze

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Igs7tm62.1(tetO-tdTomato)Hze/Igs7+involves: 129S6/SvEvTac * C57BL/6NCr

normal phenotype

References

Additional - Igs7tm62.1(tetO-tdTomato)Hze related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

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

Live Mouse

Cryorecovery - Not-For-Profit & Academic Pricing

Related Products and Services

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

Igs7^{tm62.1(tetO-tdTomato)Hze}

Allele Symbol: Igs7^{tm62.1(tetO-tdTomato)Hze}

Genotype: Igs7^{tm62.1(tetO-tdTomato)Hze}/Igs7⁺
involves: 129S6/SvEvTac * C57BL/6NCr

Additional - Igs7^{tm62.1(tetO-tdTomato)Hze} related