Overview

Also Known As: Ai6 or Ai6(RCL-ZsGreen)

Ai6 is a Cre reporter allele designed to have a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven enhanced green fluorescent protein variant (ZsGreen1) - all inserted into the Gt(ROSA)26Sor locus. Ai6 mice express robust ZsGreen1 fluorescence following Cre-mediated recombination.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation
	N5F2pN1F14 (2019-06-27 00:00:00)

Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), Reporter)	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:

$255.00 Domestic price for female 4-week

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Details

Detailed Description

Ai6 mice hemizygous for this Rosa-CAG-LSL-ZsGreen1-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette is designed to prevent high transcription levels of the downstream enhanced green fluorescent protein variant ZsGreen1 (see below). When bred to mice that express Cre recombinase, the resulting offspring will have the STOP cassette deleted in the cre-expressing tissue(s) - resulting in high expression of ZsGreen1. Because this CAG promoter-driven reporter construct is inserted into the Gt(ROSA)26Sor locus, ZsGreen1 expression is determined by which tissue(s) express Cre recombinase. These Ai6 mice are useful as a Cre reporter strain; expressing the enhanced green fluorescent protein, ZsGreen1, following Cre-mediated recombination.

Importantly, some ZsGreen1 expression may be present prior to introduction of Cre recombinase - but the ZsGreen1 expression levels after Cre recombination are significantly greater than those baseline levels. As such, it is recommended that researchers include Cre-negative Ai6 controls to establish the baseline ZsGreen1 levels in their experiments.

The donating investigator reports ZsGreen1 expression following Cre recombinase exposure can be detected by fluorescence and mRNA (in situ hybridization). Bright fluorescence is observed mainly in cell bodies.

Of note, the FRT sites flanking the mutation allow for additional targeted replacement of the reporter sequences through Flp-mediated recombination if so desired. Similarly, the attB/attP-flanked selection cassette may be removed by introduction of the site-specific bacteriophage PhiC31 integrase if so desired.

For characterization information, see images at the Allen Institute for Brain Science website (Ai6 images).

Development

The Rosa-CAG-LSL-ZsGreen1-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), ZsGreen1 sequence (from pZsGreen1-N1 (Clontech); a human codon optimized/enhanced green fluorescent protein engineered for brighter fluorescence and higher expression in mammalian cells), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. This entire construct 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 (clone Ai6) were selected and chimeric males were bred to Cre-transgenic females (expressing cre in non-germ cells from mixed genetic backgrounds). The resulting Ai6 mice (still retaining the floxed-STOP cassette and not harboring the Cre transgene) were then backcrossed to C57BL/6J for at least 1 generation prior to arrival at The Jackson Laboratory. Upon arrival, mice were further backcrossed to C57BL/6J inbred mice (Stock No. 000664) to generate this congenic colony.

Expression Data

Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	A floxed STOP prevents transcription of enhanced green fluorescent protein (ZsGreen1) until mice are bred to a Cre strain. Resulting offspring express ZsGreen1 in a pattern dictated by the Cre promoter.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Madisen L; Zwingman TA; Sunkin SM; Oh SW; Zariwala HA; Gu H; Ng LL; Palmiter RD; Hawrylycz MJ; Jones AR; Lein ES; Zeng H. 2010. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13(1):133-40PubMed: 20023653 MGI: J:155793

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Genetics

Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}

Allele Name	targeted mutation 6, Hongkui Zeng
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter)
Allele Synonym(s)	Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}; targeted mutation 6, 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
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	A floxed STOP prevents transcription of enhanced green fluorescent protein (ZsGreen1) until mice are bred to a Cre strain. Resulting offspring express ZsGreen1 in a pattern dictated by the Cre promoter.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	6
Molecular Note	The Rosa-CAG-LSL-ZsGreen1-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), ZsGreen1 sequence (from pZsGreen1-N1 (Clontech); a human codon optimized/enhanced green fluorescent protein engineered for brighter fluorescence and higher expression in mammalian cells), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus.
Mutations Made By	Hongkui Zeng, Allen Institute for Brain Science

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: cell marker for bone marrow transplantation
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Tissue/Cell Markers: Cre-lox System
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- Diabetes and Obesity Research
  - loxP
- FLP-FRT System
  - FRT-flanked Sequences
- Fluorescent Proteins
- 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
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 information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

normal phenotype

no abnormal phenotype detected
- Normal - mice are viable and fertile; used for cre reporter expression analysis

References

Madisen L; Zwingman TA; Sunkin SM; Oh SW; Zariwala HA; Gu H; Ng LL; Palmiter RD; Hawrylycz MJ; Jones AR; Lein ES; Zeng H. 2010. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13(1):133-40PubMed: 20023653 MGI: J:155793

Additional - Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Standard PCR: Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)}
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, homozygous mice may be bred.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the Ai6 or Ai6(RCL-ZsGreen) mouse strain in a publication, please cite the originating article(s) and include JAX stock #007906 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX12 (Maximum)

Pricing & Availability

Available Now

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Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service	Genotype	Price
	Heterozygous for Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)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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Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.

The Jackson Laboratory's Genotype Promise

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: Ai6 or Ai6(RCL-ZsGreen)

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze

Allele Symbol: Gt(ROSA)26Sortm6(CAG-ZsGreen1)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)26Sortm6(CAG-ZsGreen1)Hze/Gt(ROSA)26Sor+involves: 129S6/SvEvTac * C57BL/6NCr

normal phenotype

References

Additional - Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

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^{tm6(CAG-ZsGreen1)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}

Genotype: Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

Additional - Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze} related