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| 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. | |||||||||||||||||||
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Description
Strain Information
Former Names B6.Cg-Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J (Changed: 04-MAY-12 ) Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System Homozygote x Homozygote (Female x Male) 11-APR-13 Species laboratory mouse Generation N4+
Generation DefinitionsDonating Investigator Hans Clevers, Hubrecht Institute 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).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-Neor-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. Upon arrival the mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony, and then backcrossed to the same for at least 5 generations.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. 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 (mYFPA206K) 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 (ECFPS72A/Y145A/H148D/A206K) with amino acid substitutions designed to improve spectral properties and prevent dimerization.
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (358 strains)
013731 STOCK Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J
Phenotype
Phenotype Information
assigned by genotype
The following phenotype information may relate to a genetic background differing from this JAX® Mice strain.
Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/Gt(ROSA)26Sor+
involves: 129P2/OlaHsd
- no phenotypic analysis
- *normal* no phenotypic analysis (MGI Ref ID J:164644)
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
Cardiovascular Research
Cre-lox System
Cre-lox System
loxP-flanked Sequences
loxP-flanked Sequences: Test/Reporter
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: cell marker for bone marrow transplantation
Tissue/Cell Markers: spermatogonial transplantation marker
Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
Immunology and Inflammation Research
T cell deficiency, xenograft/transplant host
Neurobiology Research
cell marker
Reproductive Biology Research
Cre-lox System
spermatogonial transplantation marker
transplantation marker for embryonic and adult tissue
Toxicology Research
B and T cell deficiency, xenograft transplant host
xenograft/transplant host
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle | ||
|---|---|---|---|
| Allele Name | targeted mutation 1, Hans Clevers | ||
| Allele Type | Targeted (Reporter) | ||
| Common Name(s) | R26R-Brainbow2.1; R26R-Confetti; | ||
| Mutation Made By | Hans Clevers, Hubrecht Institute | ||
| Strain of Origin | 129P2/OlaHsd | ||
| Site of Expression | Cre recombination results in stochastic multicolor fluorescent proteins being expressed from a single genomic locus in cre-expressing tissues. | ||
| Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano | ||
| Chromosome | 6 | ||
| Gene Common Name(s) | AV258896; Gtrgeo26; Gtrosa26; R26; ROSA26; beta geo; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; | ||
| 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. [MGI Ref ID J:125961] [MGI Ref ID J:164644] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
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):134-44. [PubMed: 20887898] [MGI Ref ID J:164644]
Additional References
Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle relatedChen H; Matsumoto K; Brockway BL; Rackley CR; Liang J; Lee JH; Jiang D; Noble PW; Randell SH; Kim CF; Stripp BR. 2012. Airway epithelial progenitors are region specific and show differential responses to bleomycin-induced lung injury. Stem Cells 30(9):1948-60. [PubMed: 22696116] [MGI Ref ID J:194655]
Livet J; Weissman TA; Kang H; Draft RW; Lu J; Bennis RA; Sanes JR; Lichtman JW. 2007. Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature 450(7166):56-62. [PubMed: 17972876] [MGI Ref ID J:125961]
Sato T; van Es JH; Snippert HJ; Stange DE; Vries RG; van den Born M; Barker N; Shroyer NF; van de Wetering M; Clevers H. 2011. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature 469(7330):415-8. [PubMed: 21113151] [MGI Ref ID J:168728]
Schepers AG; Snippert HJ; Stange DE; van den Born M; van Es JH; van de Wetering M; Clevers H. 2012. Lineage tracing reveals Lgr5+ stem cell activity in mouse intestinal adenomas. Science 337(6095):730-5. [PubMed: 22855427] [MGI Ref ID J:186630]
Wang L; Benedito R; Bixel MG; Zeuschner D; Stehling M; Savendahl L; Haigh JJ; Snippert H; Clevers H; Breier G; Kiefer F; Adams RH. 2013. Identification of a clonally expanding haematopoietic compartment in bone marrow. EMBO J 32(2):219-30. [PubMed: 23188081] [MGI Ref ID J:193124]
Yan KS; Chia LA; Li X; Ootani A; Su J; Lee JY; Su N; Luo Y; Heilshorn SC; Amieva MR; Sangiorgi E; Capecchi MR; Kuo CJ. 2012. The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations. Proc Natl Acad Sci U S A 109(2):466-71. [PubMed: 22190486] [MGI Ref ID J:179994]
van Es JH; Sato T; van de Wetering M; Lyubimova A; Nee AN; Gregorieff A; Sasaki N; Zeinstra L; van den Born M; Korving J; Martens AC; Barker N; van Oudenaarden A; Clevers H. 2012. Dll1+ secretory progenitor cells revert to stem cells upon crypt damage. Nat Cell Biol 14(10):1099-104. [PubMed: 23000963] [MGI Ref ID J:193937]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, heterozygous mice may be bred together or with wildtype mice from the colony. The donating investigator reports breeding homozygous mice together with no fertility or viability problems. Mating System Homozygote x Homozygote (Female x Male) 11-APR-13 Diet Information LabDiet® 5K52/5K67
Pricing and Purchasing
Pricing, Supply Level & Notes, Controls
| Pricing for USA, Canada and Mexico shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $232.00 Female or Male Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle
Price per Pair (US dollars $) Pair Genotype $464.00 Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle x Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
| Pricing for International shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $301.60 Female or Male Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle
Price per Pair (US dollars $) Pair Genotype $603.20 Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle x Homozygous for Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
|
|
|
Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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).
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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.Ordering Information
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