LSL-miR-182 mice harbor a targeted mutation of the Gt(ROSA)26Sor locus with a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven microRNA-182 cDNA. These mice are useful for studying tumor metastasis in the lungs.
David Kirsch, Duke University Medical Center
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Inserted expressed sequence) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
LSL-miR-182 mice homozygous for this Rosa-CAG-LSL-miR-182 conditional allele are viable and fertile. A loxP-flanked STOP cassette prevents transcription of the downstream miR-182 sequence. Because this CAG promoter driven reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, miR-182 over-expression is determined by which tissue(s) express Cre recombinase. When bred to mice that express Cre recombinase, the resulting offspring will have the loxP-flanked STOP cassette deleted in the cre-expressing tissue(s). miR-182 is a microRNA markedly overexpressed in some tumors that metastasize to the lungs.
When bred to KPY mice ((mix of two floxed mice: LSL-K-rasG12D (Stock No. 008179) and p53LoxP (Stock No. 008462) with an additional LSL-YFP allele (Stock No. 006148)), mice overexpressing miR-182 show an increased rate of lung metastasis from 40% in KPY control mice to 75% in experimental mice. Also, circulating tumor cells increased.
A targeting vector was designed by Dr David Kirsch (Duke University) with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a frt site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), miR-182 sequence, 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 129Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with C57BL/6 females. The resulting LSL-miR-182 mice were maintained on a mixed background. Upon arrival, 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 | Mir182, microRNA 182, mouse, laboratory |
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Site of Expression | Mir182 is expressed in cre-expressing tissues after breeding with a strain carrying cre recombinase. |
Allele Name | targeted mutation 1, David Kirsch |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Inserted expressed sequence) |
Allele Synonym(s) | LSL-miR-182 |
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 | Mir182, microRNA 182, mouse, laboratory |
Site of Expression | Mir182 is expressed in cre-expressing tissues after breeding with a strain carrying cre recombinase. |
Strain of Origin | 129/Sv |
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), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), miR-182 sequence, 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. |
When maintaining a live colony, homozygous mice may be bred together.
When using the LSL-miR-182 mouse strain in a publication, please cite the originating article(s) and include JAX stock #028611 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm1(CAG-Mir182)Dgk> |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm1(CAG-Mir182)Dgk>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm1(CAG-Mir182)Dgk>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm1(CAG-Mir182)Dgk>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm1(CAG-Mir182)Dgk>/J Frozen Embryo | $3373.50 |
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