Overview

Also Known As:R26R-Confetti , R26R-Brainbow2.1 , Rosa26-CAG-Brainbow2.1/Confetti

The R26R-Confetti conditional allele has a CAG promoter followed by a floxed-STOP cassette and the Brainbow 2.1 construct all targeted into the Gt(ROSA)26Sor locus. The R26R-Confetti allele functions as a stochastic multicolor Cre recombinase reporter of multiple fluorescent proteins from a single genomic locus. These R26R-Confetti mice allow a way to label and distinguish individual / adjacent cells with nuclear localized, membrane-targeted, or cytoplasmic fluorescent proteins in cre recombined cells.

Donating Investigator

Hans Clevers, Hubrecht Institute

Genetic overview

Genetic Background	Generation
	N5+N1F9 (2019-08-11 00:00:00)

Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}

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
Cancer Research

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Base Price

Starting at:

$255.00 Domestic price for female 4-week

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Details

Important Note

The R26R-Confetti conditional mice are available on a C57BL/6J congenic background (Stock No. 017492), as well as a mixed genetic background (Stock No. 013731).

Detailed Description

Mice homozygous for the R26R-Confetti conditional allele are viable and fertile, with a CAG promoter, loxP site, and STOP cassette preventing transcription of the downstream Brainbow 2.1 sequences. The Brainbow 2.1 region contains two loxP-flanked dimers, each uniquely positioned in head-to-tail tandem. One dimer has nuclear-localized green fluorescent protein (hrGFPII) and a reverse-oriented cytoplasmic yellow fluorescent protein (mYFP). The other dimer has cytoplasmic red fluorescent protein (tdimer2(12)) and a reverse-oriented membrane-tethered cyan fluorescent protein (mCerulean). The Brainbow2.1 region may be written as loxP-STOP-loxP-GFP-PFY-Pxol-loxP-RFP-PFC-Pxol to show the transcriptional direction of each part. When bred to mice that express Cre recombinase, the resulting offspring may have a recombination event that stochastically places one of the four fluorescent proteins into position directly downstream of the CAG promoter within the cre-expressing tissues. Because this CAG promoter-driven Brainbow 2.1 reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, fluorescent protein expression is determined by which tissues express Cre recombinase. The donating investigator reports that mice do not express any fluorescent cells prior to introduction of Cre recombinase. The donating investigator confirms fluorescent protein expression following exposure to cre can be detected by direct fluorescence (and presumably also via mRNA (in situ hybridization) and antibody staining (immunohistochemistry)).

Initial Cre recombination outcomes may recombine the loxP-flanked STOP cassette (green), invert the loxP-STOP-loxP-GFP-PFY-Pxol region (yellow), recombine the loxP-STOP-loxP-GFP-PFY-Pxol-loxP region (red), or invert the entire loxP-STOP-loxP-GFP-PFY-Pxol-loxP-RFP-PFC-Pxol region (blue). Other recombination outcomes may not remove the STOP cassette and result in no fluorescent reporter labeling in cre-expressing cells. In addition, sequential recombination outcomes may reduce the construct to a single invertible dimer segment that can continue to invert as long as Cre recombinase is present. The donating investigator also reports that weaker cre expression favors inverting the loxP-STOP-loxP-GFP-PFY-Pxol region rather than removal of the loxP-STOP-loxP region: this results in less green-fluorescing cells / more non-fluorescing cells than is expected if using a strong cre-expressing line.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for the strain above. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

A targeting vector containing (from 5' to 3') a strong CAGG promoter (CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter), a loxP site, a PGK-Neo^r-pA cassette (serving as a transcriptional roadblock), and the Brainbow 2.1 construct (described in greater detail below). This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation into 129P2/OlaHsd-derived IB10/E14IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6 females to generate the R26R-Confetti colony. The resulting R26R-Confetti colony were bred with other mutant or cre-expressing mice, but these other mutations were bred away from the R26R-Confetti colony. The R26R-Confetti mice are reported to be on a genetic background equivalent to ~2-3 backcross generations to C57BL/6 prior to sending to The Jackson Laboratory Repository in 2010. Upon arrival, the mice were bred with C57BL/6J inbred mice (Stock No. 000664) for one generation to establish Stock No. 013731. Later, some mice were bred to C57BL/6J inbred mice for three additional generations using a marker-assisted, speed congenic approach to generate this C57BL/6J-congenic strain (Stock No. 017492).

The Brainbow 2.1 construct was designed by Drs. Jeff Lichtman and Joshua Sanes (Harvard University) with four fluorescent protein sequences uniquely positioned in a tandem fashion and delimited by loxP sites in opposite orientation. Specifically, this Brainbow 2.1 coding region is composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation, and a loxP site and monomeric EYFP (mYFP^A206K plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2(12) RFP plus polyA sequence in forward orientation, and a loxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the Brainbow 2.1 construct. The entire construct may be written as loxP-STOP-loxP-GFP-PFY-Pxol-loxP-RFP-PFC-Pxol-frt to show transcriptional direction of each part.

The hrGFPII variant of GFP (from Stratagene vector phrGFPII-C) has amino acid substitutions designed to improve spectral properties and performance in mammalian systems. The monomeric EYFP (mYFP^A206K) has an amino acid substitution replacing a hydrophobic region with a positively charged residue designed to prevent dimerization. The tdimer2(12) RFP is a non-oligomerizing DsRed variant with a 12 residue linker fusing two copies of the protein (tandem dimer). The monomeric Cerulean (mCerulean) is a variant of ECFP (ECFP^{S72A/Y145A/H148D/A206K}) with amino acid substitutions designed to improve spectral properties and prevent dimerization.

Expression Data

Expressed Gene	CFP, Cyan Fluorescent Protein, jellyfish
Expressed Gene	GFP, Green Fluorescent Protein,
Expressed Gene	YFP, Yellow Fluorescent Protein, jellyfish
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Cre recombination results in stochastic multicolor fluorescent proteins being expressed from a single genomic locus in cre-expressing tissues.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Snippert HJ; van der Flier LG; Sato T; van Es JH; van den Born M; Kroon-Veenboer C; Barker N; Klein AM; van Rheenen J; Simons BD; Clevers H. 2010. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143(1):134-44PubMed: 20887898 MGI: J:164644

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Genetics

Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}

Allele Symbol: Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}

Allele Name	targeted mutation 1, Hans Clevers
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter)
Allele Synonym(s)	R26R-Brainbow2.1; R26R-Confetti
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	CFP, Cyan Fluorescent Protein, jellyfish
Expressed Gene	GFP, Green Fluorescent Protein,
Expressed Gene	YFP, Yellow Fluorescent Protein, jellyfish
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Cre recombination results in stochastic multicolor fluorescent proteins being expressed from a single genomic locus in cre-expressing tissues.
Strain of Origin	129P2/OlaHsd
Chromosome	6
Molecular Note	A targeting vector containing (from 5' to 3') a strong CAGG promoter, a loxP site, a PGK-Neo-pA cassette (serving as a transcriptional roadblock), and the Brainbow 2.1 construct. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The Brainbow 2.1 construct was designed by Drs. Jeff Lichtman and Joshua Sanes (Harvard University) with four fluorescent protein sequences uniquely positioned in a tandem fashion and delimited by loxP sites in opposite orientation. Specifically, this Brainbow 2.1 coding region is composed of two adjacent floxed head-to-tail tandem dimers. The first head-to-tail dimer contains a loxP site and humanized Renilla GFP (hrGFPII; with nuclear localization signal plus polyA sequence) in forward orientation and a loxP site and monomeric EYFP (mYFPA206K plus polyA sequence) in reverse orientation. The second head-to-tail dimer contains a loxP site and tdimer2(12) RFP plus polyA sequence in forward orientation, and a loxP site and mCerulean CFP (with membrane tethering palmitoylation sequence plus polyA sequence) in reverse orientation. A single frt site is located at the 3' end of the Brainbow 2.1 construct.
Mutations Made By	Hans Clevers, Hubrecht Institute

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Genetics Research
  - Tissue/Cell Markers: multiple
  - Tissue/Cell Markers: cell marker for bone marrow transplantation
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Tissue/Cell Markers: spermatogonial transplantation marker
  - Tissue/Cell Markers
  - Mutagenesis and Transgenesis: Cre-lox System
  - Mutagenesis and Transgenesis
  - Tissue/Cell Markers: Cre-lox System
- Reproductive Biology Research
  - transplantation marker for embryonic and adult tissue
  - spermatogonial transplantation marker
  - Cre-lox System
- Toxicology Research
  - B and T cell deficiency, xenograft transplant host
  - xenograft/transplant host
- Fluorescent Proteins
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- Developmental Biology Research
  - transplantation marker for embryonic and adult tissue
  - Cre-lox System
- Immunology, Inflammation and Autoimmunity Research
  - T cell deficiency, xenograft/transplant host
- Neurobiology Research
  - cell marker
- Cardiovascular Research
  - Cre-lox System
- Cancer Research
  - Cre-lox System
Neurobiology Research
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- Fluorescent protein expression in neural tissue
Cancer Research
- Cre-lox System
  - loxP-flanked Sequences

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^{tm1(CAG-Brainbow2.1)Cle}/Gt(ROSA)26Sor⁺
involves: 129P2/OlaHsd

mammalian phenotype

no phenotypic analysis

References

Snippert HJ; van der Flier LG; Sato T; van Es JH; van den Born M; Kroon-Veenboer C; Barker N; Klein AM; van Rheenen J; Simons BD; Clevers H. 2010. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143(1):134-44PubMed: 20887898 MGI: J:164644

Additional - Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle} related

Technical Support

CONTACT TECHNICAL SUPPORT

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

Both heterozygous and homozygous mice are viable and fertile.
When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664).
Alternatively, homozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the R26R-Confetti , R26R-Brainbow2.1 , Rosa26-CAG-Brainbow2.1/Confetti mouse strain in a publication, please cite the originating article(s) and include JAX stock #017492 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX10 (Standard)

Pricing & Availability

Available

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Gt(ROSA)26Sor<tm1(CAG-Brainbow2.1)Cle>

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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.

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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

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General Terms and Conditions

Questions about Terms of Use

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Licensing Information

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Also Known As:R26R-Confetti , R26R-Brainbow2.1 , Rosa26-CAG-Brainbow2.1/Confetti

Donating Investigator

Genetic overview

Research Applications

Base Price

Important Note

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle

Allele Symbol: Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/Gt(ROSA)26Sor+involves: 129P2/OlaHsd

mammalian phenotype

References

Additional - Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle related

Genotyping Protocols

Dietary Information

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

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^{tm1(CAG-Brainbow2.1)Cle}

Allele Symbol: Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}

Genotype: Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle}/Gt(ROSA)26Sor⁺
involves: 129P2/OlaHsd

Additional - Gt(ROSA)26Sor^{tm1(CAG-Brainbow2.1)Cle} related