Overview

Also Known As: Ai66(RCRL-tdT)-D or Ai66D

Ai66(RCRL-tdT)Δneo mice (also called Ai66(RCRL-tdT)-D, Ai66Δneo, or Ai66D) harbor the Rosa-CAG-RoxSTOPRox-LoxSTOPLox-tdTomato::deltaNeo conditional allele designed with both a Rox-flanked STOP cassette and a loxP-flanked STOP cassette preventing transcription of the tdTomato fluorescent protein. After removal of the flanked STOP cassettes by Dre- and Cre-recombination, tdTomato fluorescence is expected in cells/tissues expressing both Dre recombinase and Cre recombinase.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Research Tools
Neurobiology Research

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Base Price

Starting at:

$2,595.00 Domestic price Cryo Recovery

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Details

Detailed Description

Ai66(RCRL-tdT)Δneo mice (also called Ai66(RCRL-tdT)-D, Ai66Δneo, or Ai66D) harbor the Rosa-CAG-RoxSTOPRox-LoxSTOPLox-tdTomato::deltaNeo conditional allele designed with both a Rox-flanked STOP cassette and a loxP-flanked STOP cassette preventing transcription of the tdTomato fluorescent protein. Although under control of the endogenous Gt(ROSA)26Sor promoter/enhancer regions and the CAG hybrid promoter, widespread expression of tdTomato is prevented by both STOP cassettes. After removal of both flanked STOP cassettes via dre- and cre-mediated recombination, expression of tdTomato is expected in cells/tissues where the expression patterns of the individual promoters driving Dre recombinase and Cre recombinase overlap.

Specifically, the donating investigator reports that Ai66(RCRL-tdT)Δneo mice have
tdTomato fluorescence in cells expressing both Dre and Cre recombinase (via breeding to Dre and Cre recombinase-expressing mice or via recombinant adeno-associated virus injections [AAV-synapsin promoter-Cre and AAV-synapsin promoter-Dre expression plasmids]). tdTomato expression is presumably also detectable by mRNA [in situ hybridization] and antibody staining/immunohistochemistry. No significant tdTomato expression is observed prior to introduction of both Dre recombinase or Cre recombinase. Extremely rare fluorescence was reported in some Cre+/Dre-/Ai66(RCRL-tdT)Δneo mice (1-3 cells within the entire cortex). The donating investigator also reports that order in which the flanked STOP cassettes are removal is arbitrary. 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 (April 2013).

For characterization information, see images at the Allen Institute for Brain Science website (Ai66(RCRL-tdT) images).

Development

These Ai66(RCRL-tdT)Δneo mice (also called Ai66(RCRL-tdT)-D, Ai66Δneo, or Ai66D) have both a Rox-flanked STOP cassette and a loxP-flanked STOP cassette upstream of the tdTomato fluorescent protein, all inserted into the Gt(ROSA)26Sor locus. The specific details are below.

The Rosa-pCAG-RSR-LSL-tdTomato-WPRE-bGHpA-AttB-PGK-Neo-polyA-AttP targeting vector was designed with (from 5' to 3')
a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS),
an Rox-flanked STOP cassette (with a short open reading frame, followed by translational stops in all three reading frames, a synthetic polyA sequence, a polyA sequence from the human growth hormone gene, and a polyA sequence from the Herpes Simplex Virus TK gene),
a loxP-flanked STOP cassette (with stop codons in all three reading frames linked to three repeats of the SV40 polyA signal),
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,
an AttB site,
a PGK promoter-Neomycin resistance gene-PGK polyA cassette,
and an AttP site.

This entire targeting vector was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation of (129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were selected.

Chimeric mice were bred to PhiC31-expressing mice (C57BL/6J congenic background; see Stock No. 007743) to remove the AttB/AttP-flanked PGK-Neo-polyA cassette and replace it with the recombined AttB/AttP site (AttL). The resulting Ai66(RCRL-tdT)Δneo mice were bred with C57BL/6J wildtype mice for at least two additional generations (and the PhiC31 gene was removed) prior to sending to The Jackson Laboratory Repository in 2013. Upon arrival, males were used to cryopreserve sperm. To establish the living Ai66(RCRL-tdT)Δneo mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

Control Suggestions

Wild-type from the colony

Approximate Controls

101045 B6129SF2/J

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

Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}

Allele Name	targeted mutation 66.1, Hongkui Zeng
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}; targeted mutation 66.1, Hongkui Zeng
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)	beta geo; ROSA26; R26; Gtrgeo26; Gtrosa26; Gtrgeo26; Gtrosa26; AV258896; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; SETD5-AS1; Thumpd3as1
Site of Expression	Expression of tdTomato fluorescence depends upon promoter used that drives both FLP and Cre recombinase.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	6
Molecular Note	The Rosa-pCAG-RSR-LSL-tdTomato-WPRE-bGHpA-AttB-PGK-Neo-polyA-AttP targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), an Rox-flanked STOP cassette (with a short open reading frame, followed by translational stops in all three reading frames, a synthetic polyA sequence, a polyA sequence from the human growth hormone gene, and a polyA sequence from the Herpes Simplex Virus TK gene), a loxP-flanked STOP cassette (with stop codons in all three reading frames linked to three repeats of the SV40 polyA signal), 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, an AttB site, a PGK promoter-Neomycin resistance gene-PGK polyA cassette, and an AttP site. PhiC31-mediated recombination removed the AttB/AttP-flanked PGK-Neo-polyA cassette and replace it with the recombined AttB/AttP site (AttL).

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Research Tools
- Genetics Research
  - Mutagenesis and Transgenesis
  - Tissue/Cell Markers: Dre-Rox System
  - Mutagenesis and Transgenesis: Dre-Rox System
  - Tissue/Cell Markers: cell marker for bone marrow transplantation
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Tissue/Cell Markers
  - Tissue/Cell Markers: Cre-lox System
  - Mutagenesis and Transgenesis: Cre-lox System
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- Dre-Rox System
  - Rox-flanked Sequences
  - Rox-flanked Sequences: Test/Reporter
- Reproductive Biology Research
  - transplantation marker for embryonic and adult tissue
  - Cre-lox System
- Diabetes and Obesity Research
  - loxP
- Fluorescent Proteins
- Developmental Biology Research
  - Cre-lox System
  - transplantation marker for embryonic and adult tissue
Neurobiology Research
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- Fluorescent protein expression in neural tissue

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}/Gt(ROSA)26Sor⁺
involves: 129S4/SvJaeSor * 129S6/SvEvTac * C57BL/6J * 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 - Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- MELT: Gt(ROSA)26Sor(tdTomato-WPRE)
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony. The donating investigator has not attempted to generate homozygous mice to date (April 2013).
- Additional Breeding and Husbandry Support
Citation
When using the Ai66(RCRL-tdT)-D or Ai66D mouse strain in a publication, please cite the originating article(s) and include JAX stock #021876 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 Gt(ROSA)26Sor<tm66.1(CAG-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.

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Frozen Mouse Embryo

$2,595.00 per straw or vial

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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.

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Also Known As: Ai66(RCRL-tdT)-D or Ai66D

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm66.1(CAG-tdTomato)Hze

Allele Symbol: Gt(ROSA)26Sortm66.1(CAG-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: Gt(ROSA)26Sortm66.1(CAG-tdTomato)Hze/Gt(ROSA)26Sor+involves: 129S4/SvJaeSor * 129S6/SvEvTac * C57BL/6J * C57BL/6NCr

normal phenotype

References

Additional - Gt(ROSA)26Sortm66.1(CAG-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

Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}

Genotype: Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze}/Gt(ROSA)26Sor⁺
involves: 129S4/SvJaeSor * 129S6/SvEvTac * C57BL/6J * C57BL/6NCr

Additional - Gt(ROSA)26Sor^{tm66.1(CAG-tdTomato)Hze} related