These Sun1-tagged mice are used in the INTACT (isolation of nuclei tagged in specific cell types) method for immunopurification of nuclei. Cre-dependent removal of a floxed STOP cassette allows expression of the SUN1 fusion protein at the inner nuclear membrane in targeted cell types. These nuclei can be immunopurified with antibodies against GFP or MYC for transcriptional and epigenetic studies.
Jeremy Nathans, Johns Hopkins University
Genetic Background | Generation |
---|---|
N1+pN1F13
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
These Sun1-tagged mice are used in the INTACT (isolation of nuclei tagged in specific cell types) method for immunopurification of nuclei. These mice contain a CAG promoter driving expression of coding sequences for the mouse nuclear membrane protein SUN1 (Sad1 and UNC84 domain containing 1) fused at its C-terminus to 2 copies of superfolder GFP (sfGFP) followed by 6 copies of Myc, inserted in the Gt(ROSA)26Sor locus. Expression of Sun1-sfGFP-Myc is prevented by an upstream loxP-flanked stop cassette. Cre-dependent removal of the loxP-STOP-loxP cassette allows expression of the SUN1 fusion protein at the inner nuclear membrane in targeted cell types. These nuclei can be immunopurified with antibodies against GFP or MYC for transcriptional and epigenetic studies. Mice homozygous for this allele are viable and fertile.
A targeting vector containing (from 5' to 3') a CAG promoter, a loxP-flanked STOP cassette, a mouse Sun1 fused with 2 copies of superfolder GFP and 6 copies of Myc, and an FRT-PGKneo-Frt cassette was inserted into the Gt(ROSA)26Sor gene. The construct was electroporated into 129-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. Resulting chimeric males were crossed to C57BL/6 mice and then intercrossed to homozygosity.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Expressed Gene | Myc, myelocytomatosis oncogene, mouse, laboratory |
Expressed Gene | Sun1, Sad1 and UNC84 domain containing 1, mouse, laboratory |
Site of Expression | The SUN1 fusion protein (SUN1 fused to sfGFP) is expressed at the inner nuclear membrane in targeted cell types where promoters driving Cre recombinase are expressed. |
Allele Name | targeted mutation 5, Jeremy Nathans |
---|---|
Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence) |
Allele Synonym(s) | R26-CAG-LSL-Sun1-sfGFP-Myc; SUN1-2xsfGFP-6xMYC |
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 | Myc, myelocytomatosis oncogene, mouse, laboratory |
Expressed Gene | Sun1, Sad1 and UNC84 domain containing 1, mouse, laboratory |
Site of Expression | The SUN1 fusion protein (SUN1 fused to sfGFP) is expressed at the inner nuclear membrane in targeted cell types where promoters driving Cre recombinase are expressed. |
Strain of Origin | 129 |
Chromosome | 6 |
Molecular Note | A targeting vector containing (from 5' to 3') a CAG promoter, a loxP site flanked STOP cassette, the mouse Sun1 cDNA fused with 2 copies of the superfolder GFP (sfGFP) green fluorescent protein gene and 6 copies of the Myc tag, and an FRT site flanked neomycin resistance gene cassette was inserted into the locus. Removal of the floxed stop cassette is required to achieve expression. |
Homozygotes and heterozygotes are viable and fertile.
When using the B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #021039 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm1(CAG-Sun1-sfGFP-Myc)Nat> |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm5(CAG-Sun1/sfGFP)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm5(CAG-Sun1/sfGFP)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm5(CAG-Sun1/sfGFP)Nat>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129-Gt(ROSA)26Sor<tm5(CAG-Sun1/sfGFP)Nat>/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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