R26-LSL-tAGO2 mice are used in the miRAP (miRNA tagging and Affinity Purification) method in conjunction with Cre recombinase expressing mice, for genetically targeted microRNA tagging and generating tissue specific miRNA expression profiles.
Z. Josh Huang, Cold Spring Harbor Laboratory
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
These mice contain a GFP and myc tagged Eif2c2 gene (tAgo2) under the control of the CAG (cytomegalovirus beta-actin enhancer) promoter, inserted in the Gt(ROSA)26Sor locus. Expression of the GFP/myc tagged Eif2c2 (tAgo2) gene is blocked by a loxP flanked STOP fragment placed between the Eif2c2 sequence and the CAG promoter. The Argonaute protein AGO2 (EIF2C2) binds to miRNAs and target mRNAs, as part of the RNA-induced silencing complex (RISC). The tAgo2 fusion gene was constructed by fusing GFP and myc tags to the N-terminus of Eif2c2. When in the presence of Cre recombinase activity, the GFP/myc tagged Eif2c2 gene is expressed, with GFP expression indicating expression of Eif2c2 and the myc epitope tag allowing affinity purification of associated miRNAs. When crossed to a strain expressing Cre recombinase, miRNAs in cre-expressing cells can be co-immunoprecipitated with tAGO2 by using a MYC antibody. miRNAs from all cells in the tissue sample can be co-immunoprecipitated with AGO2 or tAGO2 by using an AGO2 antibody. Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.
A targeting vector containing loxP site flanked STOP cassette, GFP and myc tagged Eif2c2 coding sequence, under the control of the CAG promoter was inserted in the Gt(ROSA)26Sor locus. The construct was electroporated into C57BL/6 derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to albino C57BL/6 mice. Heterozygotes were crossed to generate homozygotes. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | Ago2, argonaute RISC catalytic subunit 2, mouse, laboratory |
Expressed Gene | myc tag, myc tag, |
Site of Expression | When bred to mice that express Cre recombinase, EGFP will be expressed in tissues where promoters driving Cre recombinase are expressed. |
Allele Name | targeted mutation 1, Z Josh Huang |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) |
Allele Synonym(s) | Gt(ROSA)26Sortm1(CAG-xstpx-GFP/myc/Eif2c2)Zjh |
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 | Ago2, argonaute RISC catalytic subunit 2, mouse, laboratory |
Expressed Gene | myc tag, myc tag, |
Site of Expression | When bred to mice that express Cre recombinase, EGFP will be expressed in tissues where promoters driving Cre recombinase are expressed. |
Strain of Origin | B6.Cg-Thy1a |
Chromosome | 6 |
Molecular Note | A targeting vector containing loxP site flanked STOP cassette, GFP, and myc-tagged Eif2c2 coding sequence under the control of the CAG promoter was inserted in the Gt(ROSA)26Sor locus. When in the presence of Cre recombinase activity, the GFP/myc tagged Eif2c2 gene is expressed, GFP expression indicates expression of Eif2c2, and the myc epitope tag allowing affinity purification of associated miRNAs. |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the R26-LSL-tAGO2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #017626 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Gt(ROSA)26Sor<tm1(CAG-GFP/Eif2c2)Zjh> |
Frozen Mouse Embryo | B6(Cg)-Gt(ROSA)26Sor<tm1(CAG-GFP/Eif2c2)Zjh>/J | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Gt(ROSA)26Sor<tm1(CAG-GFP/Eif2c2)Zjh>/J | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Gt(ROSA)26Sor<tm1(CAG-GFP/Eif2c2)Zjh>/J | $3373.50 |
Frozen Mouse Embryo | B6(Cg)-Gt(ROSA)26Sor<tm1(CAG-GFP/Eif2c2)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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