Overview

Also Known As: Ai38 or Ai38(RCL-GCaMP3)

Estimated Removal of Live Colony date: 15 March 2018.

A C57BL/6J congenic version of this Ai38 strain will remain available as Stock No. 029043).

Ai38 mice express the fluorescent calcium indicator protein GCaMP3 after exposure to Cre recombinase. Following subsequent calcium binding (such as neuronal activation), bright EGFP fluorescence is observed.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)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

Neurobiology Research
Cell Biology 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

Ai38 mice heterozygous for the Rosa-CAG-LSL-GCaMP3-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette prevents transcription of the downstream GCaMP3 fusion gene (see below for detailed description of GCaMP3). Because the CAG promoter driven reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, GCaMP3 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 fluorescent calcium indicator protein, GCaMP3. The donating investigator reports that Ai38 mice do not express GCaMP3 prior to introduction of Cre recombinase. Following exposure to Cre recombinase, GCaMP3 expression (EGFP fluorescence) is detected in the cre-expressing tissues. In the absence of calcium binding, low EGFP fluorescence is reported. Following calcium binding (such as neuronal activation) bright EGFP fluorescence is observed. GCaMP3 expression in tissues other than brain has not yet been evaluated by the donating investigator (April 2011). The phenotype of homozygous mice has not been characterized by the donating investigator.

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 (Ai38 images).

GCaMP3 is a fluorescent calcium indicator. The GCaMP3 fusion gene contains the chicken smooth muscle M13 fragment of myosin light chain kinase (MYLK), a circularly permutated EGFP sequence (with several amino acid substitutions designed to increase dynamic range and baseline fluorescence), and a rat calmodulin (Calm) sequence modified to increase the fluorescence change for small calcium transients. In the absence of calcium binding, dim EGFP fluorescence is expected as there is a pore from the outside of its barrel into the chromophore. Upon calcium binding, this pore becomes occupied and fluorescence is increased.

Development

The Rosa-CAG-LSL-GCaMP3-WPRE targeting vector was designed by Dr. Hongkui Zeng (Allen Institute for Brain Science) to have (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 GCaMP3 fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette.

The GCaMP3 fusion gene contains a cDNA sequence encoding the chicken smooth muscle M13 fragment of myosin light chain kinase, a circularly permutated EGFP (amino acid residues 149-238 followed by amino acid residues 1-144 (including M153K, V163A, S175G, D180Y, T203V, A206K, and V93I mutations to increase dynamic range and baseline fluorescence), and a DNA fragment encoding amino acid residues 2-148 of the rat calmodulin (modified to harbor the N60D mutation to increase the fluorescence change for small calcium transients).

The entire Rosa-CAG-LSL-GCaMP3-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 Ai38) were selected.

Chimeric mice were bred with C57BL/6 wildtype mice and/or PhiC31-expressing mice (C57BL/6J congenic background; see Stock No. 007743); but some mice retaining the attB/attP-flanked PGK-frt-Neo-polyA cassette were propagated (see more details below).

The donating investigator reported that these mutant mice were then bred with C57BL/6J wildtype mice (but see SNP note below) for approximately four additional generations prior to sending to The Jackson Laboratory Repository. Upon arrival, sperm from Ai38 males was used to fertilize oocytes from C57BL/6J inbred mice (Stock No. 000664) in order to rederive the living colony at The Jackson Laboratory Repository.

All of the mice originally sent to The Jackson Laboratory Repository, as well as all the first generation of rederived offspring genotyped as completely co-segregating for the neo cassette and GCaMP3 portion of the Ai38 construct. As such, these mice harbor the Ai38 mutation (retaining the attB/attP-flanked PGK-frt-Neo-polyA cassette) rather than the Ai38Δneo mutation.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Expression Data

Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	Cre excision of the stop signal results in expression of a circularly permutated EGFP in cre-expressing tissues.

Control Suggestions

Wild-type from the colony
000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Feng H. 2011. Direct Data Submission 2011/05/19 MGI Direct Data SubmissionMGI: J:171735
Zariwala HA; Borghuis BG; Hoogland TM; Madisen L; Tian L; De Zeeuw CI; Zeng H; Looger LL; Svoboda K; Chen TW. 2012. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo. J Neurosci 32(9):3131-41PubMed: 22378886 MGI: J:182205

View All References

Genetics

Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze}

Allele Name	targeted mutation 38, Hongkui Zeng
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter)
Allele Synonym(s)	targeted mutation 38, Hongkui Zeng; Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)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	GFP, Green Fluorescent Protein,
Site of Expression	Cre excision of the stop signal results in expression of a circularly permutated EGFP in cre-expressing tissues.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	6
Molecular Note	The Rosa-CAG-LSL-GCaMP3-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 GCaMP3 fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. The GCaMP3 fusion gene contains a cDNA sequence encoding the chicken smooth muscle M13 fragment of myosin light chain kinase, a circularly permutated EGFP (amino acid residues 149-238 followed by amino acid residues 1-144 (including M153K, V163A, S175G, D180Y, T203V, A206K, and V93I mutations to increase dynamic range and baseline fluorescence), and a DNA fragment encoding amino acid residues 2-148 of the rat calmodulin (modified to harbor the N60D mutation to increase the fluorescence change for small calcium transients).
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
- Optogenetic and Chemogenetic tools
  - Calcium sensor expressing strains
  - Cre-dependent optogenetic tools
- Channel and Transporter Defects
  - calcium
- Fluorescent protein expression in neural tissue
Cell Biology Research
- Channel and Transporter Defects
  - calcium
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
- FLP-FRT System
  - FRT-flanked Sequences: Test/Reporter
  - FRT-flanked Sequences
- PhiC31 System
  - attB/attP-flanked Sequences

Mammalian Phenotype Terms by Genotype

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

Genotype: Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

mammalian phenotype

no phenotypic analysis

References

Feng H. 2011. Direct Data Submission 2011/05/19 MGI Direct Data SubmissionMGI: J:171735
Zariwala HA; Borghuis BG; Hoogland TM; Madisen L; Tian L; De Zeeuw CI; Zeng H; Looger LL; Svoboda K; Chen TW. 2012. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo. J Neurosci 32(9):3131-41PubMed: 22378886 MGI: J:182205

Additional - Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Separated PCR:Gt(ROSA)26Sor^(GCaMP-WPRE)
- Genotyping resources and troubleshooting
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
Citation
When using the Ai38 or Ai38(RCL-GCaMP3) mouse strain in a publication, please cite the originating article(s) and include JAX stock #014538 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<tm38.1(CAG-GCaMP3)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;129S-Gt(ROSA)26Sor<tm38(CAG-GCaMP3)Hze>/J	$2595.00
Frozen Mouse Embryo	B6;129S-Gt(ROSA)26Sor<tm38(CAG-GCaMP3)Hze>/J	$2595.00
Frozen Mouse Embryo	B6;129S-Gt(ROSA)26Sor<tm38(CAG-GCaMP3)Hze>/J	$3373.50
Frozen Mouse Embryo	B6;129S-Gt(ROSA)26Sor<tm38(CAG-GCaMP3)Hze>/J	$3373.50

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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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Strain(s) not available to companies or for-profit entities.

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Also Known As: Ai38 or Ai38(RCL-GCaMP3)

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze

Allele Symbol: Gt(ROSA)26Sortm38(CAG-GCaMP3)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)26Sortm38(CAG-GCaMP3)Hze/Gt(ROSA)26Sor+involves: 129S6/SvEvTac * C57BL/6NCr

mammalian phenotype

References

Additional - Gt(ROSA)26Sortm38(CAG-GCaMP3)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^{tm38(CAG-GCaMP3)Hze}

Allele Symbol: Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze}

Genotype: Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze}/Gt(ROSA)26Sor⁺
involves: 129S6/SvEvTac * C57BL/6NCr

Additional - Gt(ROSA)26Sor^{tm38(CAG-GCaMP3)Hze} related