These RGBow reporter mice express one of three membrane-tethered fluorescent proteins in a random manner upon cre-mediated recombination; this allows labeling of individual cell types with distinguishable color variations in cre recombined cells.
Z. Josh Huang, Cold Spring Harbor Laboratory
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
RGBow reporter mice homozygous for this conditional fluorescent allele are viable and fertile. Mice possess three farnesylated fluorescent protein sequences uniquely flanked with pairs of incompatible Lox sites alternated to create mutually exclusive recombination events. This allows stochastic expression of multiple fluorescent proteins from a single locus. Prior to cre-mediated recombination, expression of the fluorescent proteins is restricted by a translational STOP sequence. When bred to Cre recombinase expressing mice, the resulting offspring can have one of three expression outcomes in each cell of the cre expressing tissue(s): mOrange2 (monomeric RFP), EGFP, or mKate2 (monomeric far-red RFP). A farnesylation sequence tethers the fluorophores to the membrane, allowing clear labeling of each cell.
The RGBow targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), and three farnesylated fluorescent protein sequences uniquely flanked with pairs of incompatible Lox sites. Specifically, RGBow coding region contains (from 5' to 3') a LoxP site, a Lox2272 site, a LoxN site, a transcriptional STOP signal, mOrange2 sequence (derived from a monomeric red fluorescent protein) with polyA sequence, a LoxP site, EGFP (enhanced green fluorescent protein) with polyA sequence, a Lox2272 site, mKate2 (derived from a monomeric far-red fluorescent protein). This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation of (C57BL/6 x 129S4/SvJae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were selected and chimeric males were bred to C57BL/6J females and offspring were subsequently bred together to create a homozygous colony of RGBow reporter mice. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | One of either mOrange2 (monomeric RFP), EGFP, or mKate2 (monomeric far-red RFP) is expressed in cre-expressing cells after cre recombination. |
Allele Name | targeted mutation 4, Z Josh Huang |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter) |
Allele Synonym(s) | RGBow |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | One of either mOrange2 (monomeric RFP), EGFP, or mKate2 (monomeric far-red RFP) is expressed in cre-expressing cells after cre recombination. |
Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 |
Chromosome | 6 |
Molecular Note | The targeting vector is designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), and three farnesylated fluorescent protein sequences uniquely flanked with pairs of incompatible lox sites. Specifically, the coding region contains (from 5' to 3') a loxP site, a lox2272 site, a loxN site, a transcriptional STOP signal, mOrange2 sequence (derived from a monomeric red fluorescent protein) with polyA sequence, a loxP site, EGFP (enhanced green fluorescent protein) with polyA sequence, a lox2272 site, mKate2 (derived from a monomeric far-red fluorescent protein). This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. |
When maintaining a live colony, homozygous mice may be bred together.
When using the RGBow reporter mouse strain in a publication, please cite the originating article(s) and include JAX stock #028583 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm4(CAG-mOrange2,-EGFP,-mKate2)Zjh> |
Frozen Mouse Embryo | B6;129S4-Gt(ROSA)26Sor<tm4(CAG-mOrange2 -EGFP -mKate2)Zjh>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S4-Gt(ROSA)26Sor<tm4(CAG-mOrange2 -EGFP -mKate2)Zjh>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S4-Gt(ROSA)26Sor<tm4(CAG-mOrange2 -EGFP -mKate2)Zjh>/J | $3373.50 |
Frozen Mouse Embryo | B6;129S4-Gt(ROSA)26Sor<tm4(CAG-mOrange2 -EGFP -mKate2)Zjh>/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. 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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