These Ai3 mice harbor a targeted mutation of the Gt(ROSA)26Sor locus with a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven enhanced yellow fluorescent protein (EYFP), and may be useful as a Cre reporter strain. EYFP is expressed following Cre-mediated recombination.
Hongkui Zeng, Allen Institute for Brain Science
Genetic Background | Generation |
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N4+N8F9
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Starting at:
$255.00 Domestic price for female 4-week |
333.51 Domestic price for breeder pair |
Ai3 mice hemizygous for this Rosa-CAG-LSL-EYFP-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette prevents transcription of the downstream enhanced yellow fluorescent protein (EYFP). When bred to mice that express Cre recombinase, the resulting offspring will have the STOP cassette deleted in the cre-expressing tissue(s); resulting in expression of EYFP. Because this CAG promoter driven reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, EYFP expression is determined by which tissue(s) express Cre recombinase. The donating investigator reports that Ai3 mice do not express EYFP prior to introduction of Cre recombinase and EYFP expression following exposure to cre can be detected by fluorescence, mRNA (in situ hybridization) and antibody staining (immunohistochemistry). Of note, the FRT sites flanking the mutation allow for additional targeted replacement of the reporter sequences through Flp-mediated recombination if so desired. Similarly, the attB/attP-flanked selection cassette may be removed by introduction of the site-specific bacteriophage PhiC31 integrase if so desired. These Ai3 mice are useful as a Cre reporter strain; expressing enhanced yellow fluorescent protein following Cre-mediated recombination.
For characterization information, see images at the Allen Institute for Brain Science website (Ai3 images).
The Rosa-CAG-LSL-EYFP-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), an enhanced yellow fluorescent protein sequence (from pcDNA 6.2/N-YFP-DEST (Invitrogen)), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation of (129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells (clone Ai3) were selected and chimeric males were bred to C57BL/6J females. The resulting Ai3 mice were then backcrossed to C57BL/6J for at least 3 generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were further backcrossed to C57BL/6J inbred mice (Stock No. 000664) to generate this congenic colony.
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
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Site of Expression | A floxed STOP prevents transcription of EYFP until mice are bred to a Cre strain. Resulting offspring express EYFP in a pattern dictated by the Cre promoter. |
Allele Name | targeted mutation 3, Hongkui Zeng |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter) |
Allele Synonym(s) | Ai3; R26LSL-YFP; Rosa-CAG-LSL-EYFP-WPRE |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
Site of Expression | A floxed STOP prevents transcription of EYFP until mice are bred to a Cre strain. Resulting offspring express EYFP in a pattern dictated by the Cre promoter. |
Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Chromosome | 6 |
Molecular Note | The Rosa-CAG-LSL-EYFP-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), an enhanced yellow fluorescent protein sequence (from pcDNA6.2/N-YFP-DEST), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a polyA signal, and an attB/attP-flanked PGK-FRT-Neo-polyA cassette. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. |
Mutations Made By | Hongkui Zeng, Allen Institute for Brain Science |
When maintaining a live colony, heterozygous mice may be bred with wildtype mice from the colony or to C57BL/6J inbred mice. The donating investigator has not attempted to breed homozygous mice together.
When using the Ai3 or Ai3(RCL-EYFP) mouse strain in a publication, please cite the originating article(s) and include JAX stock #007903 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm3(CAG-EYFP)Hze>/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm3(CAG-EYFP)Hze>/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm3(CAG-EYFP)Hze>/J | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm3(CAG-EYFP)Hze>/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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