These IRG transgenic mice are a double-fluorescent, Cre-reporter strain; with widespread expression of red fluorescence prior to Cre recombinase exposure, and green fluorescence following cre-mediated recombination in a pattern determined by cre expression, and should provide a versatile tool for analyzing complex cellular relationships in a wide variety of tissues.
Gregory Elder, James J. Peters VA Medical Center
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter) |
Hemizygous IRG transgenic mice are viable and fertile, with widespread expression of a loxP-flanked optimized red fluorescent protein variant (DsRed-Express) directed to embryonic and adult tissues by the CAG promoter prior to exposure to Cre recombinase. When bred to cre-expressing mice, the resulting offspring have the DsRed-Express cassette deleted in the cre-expressing tissue(s), allowing expression of the enhanced green fluorescent protein (EGFP) cassette located just downstream. These IRG transgenic mice are a double-fluorescent, Cre-reporter strain; with widespread expression of red fluorescence prior to Cre recombinase exposure, and green fluorescence following cre-mediated recombination in a pattern determined by cre expression, and should provide a versatile tool for analyzing complex cellular relationships in a wide variety of tissues.
For example, when using IRG transgenic mice along with Nestin-Cre mice (see Stock No. 003771), red and green native fluorescence can be imaged simultaneously in thick tissue sections by confocal microscopy; thus allowing for analysis of complex reconstructions suitable for studying neuronal morphologies as well as neurovascular interactions in brain.
The CCAG-R/G expression vector was designed with (from 5' to 3') a Human cytomegalovirus immediate early enhancer and chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a loxP site, a DsRed-Express protein sequence followed by three SV40 polyadenylation signals (DsRed-Express (Clontech) is a red fluorescent protein tetramer with improved solubility and fluorescence detection), a second loxP site, an enhanced green fluorescent protein (EGFP) sequence itself followed by a rabbit beta-globin polyadenylation signal. To minimize integration site effects and possible promoter interference, the CCAG-R/S expression vector was flanked on both sides with two paired copies of anti-sense oriented 5'-HS4 insulator sequences from the chicken beta-globin gene; generating the final "insulator/red/green" targeting vector (pIRG). The donating investigator confirms that this 9.4 kb transgene was injected into fertilized oocytes from B6C3F1 hybrid mice. Founder mice were bred to C57BL/6 wild-type mice to generate IRG line 5 (estimated to contain seven copies of the transgene). These IRG mice were maintained on a mixed C57BL/6 and C3H genetic background by breeding hemizygotes with C57BL/6 or B6C3 hybrid mice for many generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | Widespread expression of red fluorescent protein is observed; following cre-mediated recombination, GFP is expressed. |
Allele Name | transgene insertion 5, Gregory A Elder |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | insulator/red/green; IRG |
Gene Symbol and Name | Tg(CAG-DsRed,-EGFP)5Gae, transgene insertion 5, Gregory A Elder |
Gene Synonym(s) | |
Promoter | ACTB, actin, beta, chicken |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | Widespread expression of red fluorescent protein is observed; following cre-mediated recombination, GFP is expressed. |
Strain of Origin | (C57BL/6 x C3H)F1 |
Chromosome | UN |
Molecular Note | The CCAG-R/G expression vector was designed with (from 5' to 3') a Human cytomegalovirus immediate early enhancer and chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a loxP site, a DsRed-Express protein sequence followed by three SV40 polyadenylation signals (DsRed-Express (Clontech) is a red fluorescent protein tetramer with improved solubility and fluorescence detection), a second loxP site, an enhanced green fluorescent protein (EGFP) sequence itself followed by a rabbit beta-globin polyadenylation signal. To minimize integration site effects and possible promoter interference, the CCAG-R/G expression vector was flanked on both sides with two paired copies of anti-sense oriented 5'-HS4 insulator sequences from the chicken beta-globin gene; generating the final "insulator/red/green" targeting vector (pIRG). Line 5 was established and has an estimated transgene copy number of 7. Embryonic and adult tissues exhibit widespread red fluorescent protein expression in the absence of cre recombinase. |
Mutations Made By | Gregory Elder, James J. Peters VA Medical Center |
When maintaining a live colony, hemizygous mice may be bred with noncarrier (wildtype) mice.
When using the B6;C3-Tg(CAG-DsRed,-EGFP)5Gae/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #008605 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous or Non carrier for Tg(CAG-DsRed,-EGFP)5Gae |
Frozen Mouse Embryo | B6;C3-Tg(CAG-DsRed -EGFP)5Gae/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;C3-Tg(CAG-DsRed -EGFP)5Gae/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;C3-Tg(CAG-DsRed -EGFP)5Gae/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;C3-Tg(CAG-DsRed -EGFP)5Gae/J Frozen Embryo | $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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