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| These Brainbow 2.1 (founder line R) mice allow labeling of individual neurons in peripheral and central neurons with nuclear localized, membrane-targeted, or cytoplasmic fluorescent proteins in cre recombined cells, and may also be useful in conjunction with other Brainbow strains (Stock No. 007901, Stock No. 007910, Stock No. 007911) for neurobiological studies. | |||||||||||||||
- Description
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Description
Strain Information
Former Names B6.Cg-Tg(Thy1-hrGFPII,-mYFP,-tdimer2(12),-mCerulean)R21Lich/J (Changed: 15-NOV-07 ) Type Congenic; Mutant Strain; Transgenic; 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) 10-DEC-08 Species laboratory mouse Generation N6+F3 (06-JAN-09) Donating Investigator Jeff Lichtman, Harvard University Donating Investigator Joshua Sanes, Harvard University Description
View Brainbow images for Stock No's 007901, 007910, 007911, and 007921.
These Thy1-Brainbow 2.1 (line R) transgenic mice are viable and fertile. The mice possess two invertible DNA segments (four fluorescent protein sequences in total) uniquely positioned in tandem and flanked with LoxP sites to generate a larger number of recombination outcomes; allowing stochastic expression of multiple fluorescent proteins from a single transgene. Although the fluorescent protein immediately adjacent to the promoter, hrGFPII (with nuclear localization signal), was designed to be expressed prior to Cre-mediated recombination, basal hrGFPII expression may not be observed in mouse tissues. When bred to Cre recombinase expressing mice, however, the resulting offspring can have one of three different inversions for each transgene in each cell of the cre expressing tissue(s). In addition, two excision events may reduce the construct to one of two single invertible DNA segments which can continue to invert as long as cre is present. These different recombination possibilities can be used to express four genes (hrGFPII, mYFP, tdimer2(12) (RFP), or mCerulean (CFP)) to the peripheral and central neurons. A nuclear localization signal targets the hrGFPII to the nucleus, a palmitoylation sequence tethers the mCerulean (CFP) to the membrane (allowing clear labeling of axonal processes), and cytoplasmic tdimer2(12) (RFP) and mYFP better labeled neuronal cell bodies and dendrites. Integration of tandem transgene copies yields combinatorial fluorescent protein expression in each cell, and thus many possible cell colors, providing a way to distinguish adjacent neurons and visualize other cellular interactions. Of note, the single FRT site inserted at the 5' end of the transgene allows tandem transgene copy number reduction through Flp-mediated recombination if desired. These Brainbow 2.1 (founder line R) mice allow labeling of individual neurons in peripheral and central neurons with nuclear localized, membrane-targeted, or cytoplasmic fluorescent proteins in cre recombined cells, and may also be useful in conjunction with other Brainbow strains (Stock No. 007901, Stock No. 007910, Stock No. 007911) for neurobiological studies.Development
The Thy1-Brainbow 2.1 transgene was designed with the mouse Thy1 regulatory elements surrounding four fluorescent proteins (green (GFP), enhanced yellow (EYFP), red (RFP), and cyan (CFP)) 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 head to tail tandem dimers, the first contains a loxP site, humanized Renilla GFP (hrGFPII (Stratagene); with nuclear localization signal), and polyA sequence in forward orientation, and a LoxP site, monomeric EYFP (mYFPA206K), and polyA sequence in reverse orientation. The second head to tail dimer contains a LoxP site, tdimer2(12) RFP, and polyA sequence in forward orientation, and a LoxP site, mCerulean CFP (with membrane tethering palmitoylation sequence), and polyA sequence in reverse orientation. A single FRT site is located at the 3' end of the coding region. This transgene was microinjected into C57BL/6JxCBA hybrid oocytes. Founder mice (line R) were bred with C57BL/6. The resulting mutant mice were backcrossed to C57BL/6 for at least 5 generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.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 (225 strains)
004218 B6.129(ICR)-Tg(CAG-ECFP)CK6Nagy/J 007901 B6.Cg-Tg(Thy1-Brainbow1.0)HLich/J 007911 B6.Cg-Tg(Thy1-Brainbow1.1)MLich/J 003710 B6.Cg-Tg(Thy1-CFP)23Jrs/J 007940 B6.Cg-Tg(Thy1-CFP/COX8A)C1Lich/J 007967 B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J 007910 B6;CBA-Tg(Thy1-Brainbow1.0)LLich/J 006617 B6;CBA-Tg(Thy1-CFP/COX8A)S2Lich/J 008004 B6;SJL-Tg(Thy1-ECFP/VAMP2)1Sud/J 007856 B6SJL-Tg(Thy1-Syt1/ECFP)1Sud/J 003773 STOCK Tg(CAG-ECFP)CK6Nagy/J 006784 STOCK Tg(Ins1-ECFP)24Hara/J
005483 129-Tg(CAG-EYFP)7AC5Nagy/J 006148 B6.129X1-Gt(ROSA)26Sortm1(EYFP)Cos/J 008299 B6.Cg-Tg(NEFL-EYFP/Nefh)40Gsn/J 007901 B6.Cg-Tg(Thy1-Brainbow1.0)HLich/J 007911 B6.Cg-Tg(Thy1-Brainbow1.1)MLich/J 007612 B6.Cg-Tg(Thy1-COP4/EYFP)18Gfng/J 007615 B6.Cg-Tg(Thy1-COP4/EYFP)9Gfng/J 005630 B6.Cg-Tg(Thy1-EYFP)15Jrs/J 003709 B6.Cg-Tg(Thy1-YFP)16Jrs/J 005627 B6.Cg-Tg(Thy1-YFP/Syp)10Jrs/J 003782 B6.Cg-Tg(Thy1-YFPH)2Jrs/J 007606 B6.Cg-Tg(Thy1-cre/ESR1,-EYFP)AGfng/J 008636 B6;C-Tg(Prnp-APP695*/EYFP)49Gsn/J 007910 B6;CBA-Tg(Thy1-Brainbow1.0)LLich/J 005620 B6;D2-Tg(S100B-EYFP)1Wjt/J 007610 B6;SJL-Tg(Thy1-cre/ESR1,-EYFP)VGfng/J 007880 B6SJL-Tg(Thy1-Stx1a/EYFP)1Sud/J 006618 C57BL/6-Tg(tetO-COX8A/EYFP)1Ksn/J 006362 C57BL/6J-Tg(CMV-Cox8a/EYFP)17J/J 007857 C57BL/6J-Tg(Eno2-YFP/Cox8a)YRwb/J 007860 C57BL/6J-Tg(Eno2-YFP/Cox8a)ZRwb/J 009422 NOD.Cg-Tg(Itgax-EYFP)1Mnz/QtngJ 005130 STOCK Gt(ROSA)26Sortm1(Smo/EYFP)Amc/J
Additional Web Information
FLP-FRT Systems
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
JAX® NOTES, Winter 2008; 508. Brainbow Mice Soon to be Available From The Jackson Laboratory.
Phenotype
Phenotype Information
This mouse can be used to support research in many areas including:
CFP relatedNeurobiology Research
Cre-lox System
loxP-flanked Sequences: Test/Reporter
Fluorescent protein expression in neural tissue
Research Tools
Cre-lox System
loxP-flanked Sequences: Test/Reporter
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
FLP-FRT System
FRT-flanked Sequences
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: astrocyte-specific marker
Tissue/Cell Markers: astrocytes
Tissue/Cell Markers: astrocytes, neurons
Tissue/Cell Markers: multiple
Tissue/Cell Markers: neurons
Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
Neurobiology Research
astrocyte-specific marker
cell marker
GFP relatedResearch Tools
Fluorescent Proteins
YFP related
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(Thy1-Brainbow2.1)RLich | ||
|---|---|---|---|
| Allele Name | transgene insertion R, Jeff W Lichtman | ||
| Allele Type | Transgenic (Reporter) | ||
| Common Name(s) | Tg(Thy1-hrGFPII,-mYFP,-tdimer2(12),-mCerulean)R21Lich; Thy1-Brainbow-2.1 line R; | ||
| Mutation Made By | Jeff Lichtman and Joshua Sanes, Harvard University | ||
| Strain of Origin | C57BL/6J x CBA | ||
| Site of Expression | When bred to Cre recombinase expressing mice, there is one of three different inversions for each transgene in each cell of the cre expressing tissue(s). In addition, two excision events may reduce the construct to one of two single invertible DNA segments which can continue to invert as long as cre is present. These different recombination possibilities can be used to express four genes (hrGFPII, mYFP, tdimer2(12) (RFP), or mCerulean (CFP)). Integration of multiple tandem transgene copies allows for labeling of individual neurons and glia in peripheral and central neurons with nuclear localized, membrane-targeted, or cytoplasmic fluorescent proteins in cre recombined cells. | ||
| Expressed Gene | RFP, Red Fluorescent Protein, jelly fish | ||
| Red Fluorescent Protein (RFP), derived from marine invertebrate organisms such as the soft coral Discosoma spp and reef coral, Heteractis crispa, is a versatile reporter molecule which has found use in many biological applications. The wild type protein, which is an obligate tetramer, is not well tolerated in mammalian systems. The original molecule has been modified in order to optimize expression to mammalian physiology (examples include monomeric RFP, mRFP1, DsRed, etc). | |||
| Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish | ||
| Yellow Fluorescent Protein (YFP) is a derivative of Green Fluorescent Protein (GFP), a versatile reporter molecule which has found use in many biological applications. The original molecule has been modified in order to enhance fluorescence intensity and shift the wavelength emitted when excited. When YFP is utilized in a transgenic construct, tissue expressing sufficient amounts of YFP will fluoresce yellowish-green when exposed to a 513 nm light source. | |||
| Expressed Gene | GFP, Green Fluorescent Protein, jellyfish | ||
| Green Fluorescent Protein (GFP), derived from the jellyfish Aequorea victoria, is a versatile reporter molecule which has found use in many biological applications. In some constructs the original molecule has been modified in order to enhance its fluorescence intensity (EGFP, enhanced GFP). When utilized in a transgenic construct, tissue expressing sufficient amounts of GFP will fluoresce when exposed to a 488 nm light source. | |||
| Expressed Gene | CFP, Cyan Fluorescent Protein, jellyfish | ||
| Cyan Fluorescent Protein (CFP) is a derivative of Green Fluorescent Protein (GFP), a versatile reporter molecule which has found use in many biological applications. The original molecule has been modified in order to enhance fluorescence intensity and shift the wavelength emitted when excited. When CFP is utilized in a transgenic construct, tissue expressing sufficient amounts of CFP will fluoresce cyan when exposed to a 433 nm light source. | |||
| Promoter | Thy1, thymus cell antigen 1, theta, mouse, laboratory | ||
| Molecular Note | The Thy1-Brainbow 2.1 transgene was designed with the mouse Thy1 regulatory elements surrounding four fluorescent proteins (green (GFP), enhanced yellow (EYFP), red (RFP), and cyan (CFP)) sequences uniquely positioned in a tandem fashion and delimited byLoxP sites in opposite orientation. Specifically, this Brainbow 2.1 coding region is composed of two adjacent head to tail tandem dimers, the first contains a LoxP site, humanized Renilla GFP (hrGFPII (Stratagene); with nuclear localization signal), and polyA sequence in forward orientation, and a LoxP site, monomeric EYFP (mYFPA206K), and polyA sequence in reverse orientation. The second head to tail dimer contains a LoxP site, tdimer2(12) RFP, and polyA sequence in forward orientation, and a LoxP site,mCerulean CFP (with membrane tethering palmitoylation sequence), and polyA sequence in reverse orientation. A single FRT site is located at the 3' end of the coding region.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. [MGI Ref ID J:125961] | ||
| Gene Symbol and Name | Tg(Thy1-Brainbow2.1)RLich, transgene insertion R, Jeff W Lichtman | ||
| Chromosome | UN | ||
| Gene Common Name(s) | Tg(Thy1-hrGFPII,-mYFP,-tdimer2(12),-mCerulean)R21Lich; Thy1-Brainbow-2.1 line R; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Fluorescent Proteins (Generic GFP), Standard PCR
Fluorescent Proteins -- Generic GFP, QPCR
Tg(Thy1-Brainbow2.1)RLich, Separated PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
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]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX12
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, homozygous mice may be bred together. Mating System Homozygote x Homozygote (Female x Male) 10-DEC-08 Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $209.90 Female or Male Homozygous for Tg(Thy1-Brainbow2.1)RLich
Pairs /Price (US dollars $) Pair Genotype $419.80 Homozygous for Tg(Thy1-Brainbow2.1)RLich x Homozygous for Tg(Thy1-Brainbow2.1)RLich
Additional Supply Details
| Pricing for International shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $272.90 Female or Male Homozygous for Tg(Thy1-Brainbow2.1)RLich
Pairs /Price (US dollars $) Pair Genotype $545.80 Homozygous for Tg(Thy1-Brainbow2.1)RLich x Homozygous for Tg(Thy1-Brainbow2.1)RLich
Additional Supply Details
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Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of approximately nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within two business days following order placement. |
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| Supply Notes |
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Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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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 and Purchasing Information
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