Overview

Also Known As: Ai34D or Ai34(RCL-Syp/tdT)-D

Ai34D mice harbor a knock-in mutation of the Gt(ROSA)26Sor locus with a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven Synaptophysin-tdTomato fusion gene. Following Cre-mediated removal of the STOP cassette, Synaptophysin-tdTomato expression is observed at synaptic termini in brain regions.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation
	F?+N1F19 (2019-03-18 00:00:00)

Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Neurobiology Research
Research Tools

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Base Price

Starting at:

$278.00 Domestic price for female

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Details

Detailed Description

Ai34D (or Ai34Δneo) mice heterozygous for the Rosa-CAG-LSL-Synaptophysin-tdTomato-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette prevents transcription of the downstream Synaptophysin-tdTomato fusion gene. The Synaptophysin-tdTomato fusion protein is composed of near-full-length mouse synaptophysin protein fused in-frame to the amino terminus of the tdTomato fluorescent protein. Because the CAG promoter driven reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, Synaptophysin-tdTomato expression is determined by which tissue(s) express Cre recombinase. When bred to mice that express Cre recombinase, the resulting offspring will have the STOP cassette deleted in the cre-expressing tissues; resulting in expression of the Synaptophysin-tdTomato fusion protein. The donating investigator reports that Ai34D mice do not express Synaptophysin-tdTomato prior to introduction of Cre recombinase. Following exposure to Cre recombinase, tdTomato expression can be detected by tdTomato fluorescence and mRNA (in situ hybridization) [and presumably by antibody staining (immunohistochemistry); although this was not tested by the donating investigator]. tdTomato fluorescence is found to be concentrated at synaptic termini in brain regions. Fusion protein expression in tissues other than brain has not yet been evaluated by the donating investigator (April 2011).

Unlike the Ai34 mice from which they were derived, these Ai34D mice no longer harbor the downstream FRT site or attB/attP-flanked selection cassette. The phenotype of homozygous mice has not been characterized by the donating investigator.

For characterization information, see images at the Allen Institute for Brain Science website (Ai34 images).

Development

The Rosa-CAG-LSL-Synaptophysin-tdTomato-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), a Synaptophysin-tdTomato fusion protein sequence, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette.

To create the Synaptophysin-tdTomato fusion protein sequence, a mouse synaptophysin cDNA sequence (encoding a 308 amino acid transcript with all but the first six amino acids of the full-length protein) was fused in-frame via a six nucleotide linker to the amino terminus of the tdTomato sequence (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)).

The entire Rosa-CAG-LSL-Synaptophysin-tdTomato-WPRE 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 (clone Ai34) were selected. Chimeric mice were bred to PhiC31-expressing mice (C57BL/6J congenic background; see Stock No. 007743) to remove the PGK-FRT-Neo-polyA cassette and replace it with the recombined attB/attP site (attL). The resulting Ai34D (or Ai34Δneo) mice were bred with C57BL/6J wildtype mice for at least two more generations (and the PhiC31 gene was removed) prior to sending to The Jackson Laboratory Repository. Upon arrival, Ai34D mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generations to establish the colony.

Expression Data

Expressed Gene	RFP, Red Fluorescent Protein, coral
Expressed Gene	Syp, synaptophysin, mouse, laboratory
Site of Expression	Cre excision of the stop signal results in expression of a Synaptophysin-tdTomato fusion protein in cre-expressing tissues.

Control Suggestions

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Zeng H. 2011. Direct Data Submission 2011/04/15 MGI Direct Data SubmissionMGI: J:170755

View All References

Genetics

Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}

Allele Name	targeted mutation 34.1, Hongkui Zeng
Allele Type	Targeted (Reporter)
Allele Synonym(s)	targeted mutation 34.1, Hongkui Zeng; Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}
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
Promoter	CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter,
Expressed Gene	RFP, Red Fluorescent Protein, coral
Expressed Gene	Syp, synaptophysin, mouse, laboratory
Site of Expression	Cre excision of the stop signal results in expression of a Synaptophysin-tdTomato fusion protein in cre-expressing tissues.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	6
Molecular Note	The Rosa-CAG-LSL-Synaptophysin-tdTomato-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), a Synaptophysin-tdTomato fusion protein sequence, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. To create the Synaptophysin-tdTomato fusion protein sequence, a mouse synaptophysin cDNA sequence (encoding a 308 amino acid transcript with all but the first six amino acids of the full-length protein) was fused in-frame via a six nucleotide linker to the amino terminus of the tdTomato sequence (a non-oligomerizing DsRed fluorescent protein variant with a 12 residue linker fusing two copies of the protein (tandem dimer)). PhiC31-mediated recombination removed the neo cassette.
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:

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

Mammalian Phenotype Terms by Genotype

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

Genotype: Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

mammalian phenotype

no phenotypic analysis

References

Zeng H. 2011. Direct Data Submission 2011/04/15 MGI Direct Data SubmissionMGI: J:170755

Additional - Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- MELT: Gt(ROSA)26Sor(tdTomato-WPRE)
- Standard PCR: Gt(ROSA)26Sor(Syp)
- Genotyping resources and troubleshooting
Dietary Information
- LabDiet® 5K52 formulation (6% fat)
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred to wildtype siblings or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator has not tried to generate homozygous mice.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the Ai34D or Ai34(RCL-Syp/tdT)-D mouse strain in a publication, please cite the originating article(s) and include JAX stock #012570 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX10 (Standard)

Pricing & Availability

Available

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Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Not-For-Profit & Academic Pricing

Service	Genotype	Price
	Hemizygous or Non carrier for Gt(ROSA)26Sor<tm34.1(CAG-Syp/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.

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Also Known As: Ai34D or Ai34(RCL-Syp/tdT)-D

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm34.1(CAG-Syp/tdTomato)Hze

Allele Symbol: Gt(ROSA)26Sortm34.1(CAG-Syp/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)26Sortm34.1(CAG-Syp/tdTomato)Hze/Gt(ROSA)26Sor+involves: 129S6/SvEvTac * C57BL/6NCr

mammalian phenotype

References

Additional - Gt(ROSA)26Sortm34.1(CAG-Syp/tdTomato)Hze related

Genotyping Protocols

Dietary Information

Breeding Considerations

Mating System

Citation

Animal Health Reports

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^{tm34.1(CAG-Syp/tdTomato)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}

Genotype: Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

Additional - Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze} related