The GFP reporter inserted downstream of the Setd5 promoter eliminates Setd5 transcription (creating a null allele) and provides a reporter for Setd5 expression. SETD5 (SET domain-containing protein 5) is involved in epigenetic repression of gene expression.
Mark A. Magnuson, Vanderbilt University School of Medicine
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Setd5 | SET domain containing 5 |
Heterozygous mice harboring the Setd5.GFP reporter are viable and fertile. Mice homozygous for the mutation are not viable. Insertion of the GFP reporter downstream of the Setd5 promoter eliminates Setd5 transcription (creating a null allele) and provides a reporter for Setd5 gene expression. Setd5 is located upstream of Rosa26 and is transcribed in the opposite orientation. SETD5 (SET domain-containing protein 5) is involved in epigenetic repression of gene expression.
Using the ROSA26LCA or Loxed Cassette Acceptor allele (Gt(ROSA)26Sortm1Mgn), which contains a loxP-flanked (lox71, lox2272) selection cassette in the ROSA26 promoter, a donor cassette is inserted into the acceptor allele via recombinase mediated cassette exchange (RMCE). The donor cassette or signaling sentinel construct includes an FRT-flanked pgk-hygromycin and a GFP-SV40 polyA reporter sequence. The GFP reporter inserted into exon1 of the Setd5 gene, which is located upstream of the Rosa26 exon 1 and is transcribed in the opposite orientation. The construct was electroporated into 129S6/SvEvTac-derived TL1 ES cells. Chimeric mice were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J transgenic mice to delete the pgk-hygromycin cassette. The donating investigator reported that the mice were backcrossed to CD-1 mice for 4 generations prior to sending to MMRRC at JAX. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression |
Allele Name | targeted mutation 1.1, Mark Magnuson |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Setd5GFP |
Gene Symbol and Name | Setd5, SET domain containing 5 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | The pRosa26.Ex1.Setd5GFP exchange vector was made on a backbone of a basal Rosa26 exchange vector which contains a 5.166 kb sequence from the Rosa26 locus, Lox71/Lox2272 sites, and flrted (flanked by FRT) Pgk-Hygro selection cassette that is used for positive selection after RMCE. A GFP-SV40 polyA sequence was inserted into exon 1 of the Setd5 gene which is located upstream of the Rosa26 exon 1 and is transcribed in the opposite orientation. Gene targeting and RMCE was performed in mESCs derived from 129S6 mice. The chimeric mice were crossed with Flpe mice to remove the hygromycin resistance gene. |
While maintaining a live colony, these mice are bred as heterozygotes. Mice homozygous for the mutation are not viable.
When using the STOCK Setd5tm1.1Mgn/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36985 in your Materials and Methods section.
Facility Barrier Level Descriptions
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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