Ai6 is a Cre reporter allele designed to have a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven enhanced green fluorescent protein variant (ZsGreen1) - all inserted into the Gt(ROSA)26Sor locus. Ai6 mice express robust ZsGreen1 fluorescence following Cre-mediated recombination.
Hongkui Zeng, Allen Institute for Brain Science
Genetic Background | Generation |
---|---|
N5F2pN2F8
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Ai6 mice hemizygous for this Rosa-CAG-LSL-ZsGreen1-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette is designed to prevent high transcription levels of the downstream enhanced green fluorescent protein variant ZsGreen1 (see below).
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 high expression of ZsGreen1. Because this CAG promoter-driven reporter construct is inserted into the Gt(ROSA)26Sor locus, ZsGreen1 expression is determined by which tissue(s) express Cre recombinase. These Ai6 mice are useful as a Cre reporter strain; expressing the enhanced green fluorescent protein, ZsGreen1, following Cre-mediated recombination.
Importantly, some ZsGreen1 expression may be present prior to introduction of Cre recombinase - but the ZsGreen1 expression levels after Cre recombination are significantly greater than those baseline levels. As such, it is recommended that researchers include Cre-negative Ai6 controls to establish the baseline ZsGreen1 levels in their experiments.
The donating investigator reports ZsGreen1 expression following Cre recombinase exposure can be detected by fluorescence and mRNA (in situ hybridization). Bright fluorescence is observed mainly in cell bodies.
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.
For characterization information, see images at the Allen Institute for Brain Science website (Ai6 images).
The Rosa-CAG-LSL-ZsGreen1-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), ZsGreen1 sequence (from pZsGreen1-N1 (Clontech); a human codon optimized/enhanced green fluorescent protein engineered for brighter fluorescence and higher expression in mammalian cells), 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 Ai6) were selected and chimeric males were bred to Cre-transgenic females (expressing cre in non-germ cells from mixed genetic backgrounds). The resulting Ai6 mice (still retaining the floxed-STOP cassette and not harboring the Cre transgene) were then backcrossed to C57BL/6J for at least 1 generation 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 | GFP, Green Fluorescent Protein, |
---|---|
Site of Expression | A floxed STOP prevents transcription of enhanced green fluorescent protein (ZsGreen1) until mice are bred to a Cre strain. Resulting offspring express ZsGreen1 in a pattern dictated by the Cre promoter. |
Allele Name | targeted mutation 6, Hongkui Zeng |
---|---|
Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter) |
Allele Synonym(s) | Ai6; R26floxstop-EGFP; R26ZsGreen; RosaZsG; ROSA26LoxP-STOP-LoxP-ZsGreen1; Rosa-CAG-LSL-ZsGreen1-WPRE; Rs-ZsGreen-R; ZsGrn |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | A floxed STOP prevents transcription of enhanced green fluorescent protein (ZsGreen1) until mice are bred to a Cre strain. Resulting offspring express ZsGreen1 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-ZsGreen1-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), ZsGreen1 sequence (from pZsGreen1-N1 (Clontech); a human codon optimized/enhanced green fluorescent protein engineered for brighter fluorescence and higher expression in mammalian cells), 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, homozygous mice may be bred.
When using the Ai6 or Ai6(RCL-ZsGreen) mouse strain in a publication, please cite the originating article(s) and include JAX stock #007906 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)Hze> |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)Hze>/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)Hze>/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)Hze>/J | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm6(CAG-ZsGreen1)Hze>/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.