These mice have the Ple111-EGFP transgene targeted as a single copy "knockin" into the upstream region of the Hprt locus on the X chromosome. The promoter/regulatory regions of the human HCRT gene direct expression of EGFP.
Elizabeth M Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Hprt | hypoxanthine guanine phosphoribosyl transferase |
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | EGFP is expressed in a small cluster of neurons in the hypothalamus. |
Allele Name | targeted mutation 15, Elizabeth M Simpson |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | HCRT-EGFP; Hprt1tm15(mEMS1774)Ems; Hprt1tm15(Ple111-EGFP;mEMS1774)Ems; Ple111 |
Gene Symbol and Name | Hprt, hypoxanthine guanine phosphoribosyl transferase |
Gene Synonym(s) | |
Promoter | HCRT, hypocretin neuropeptide precursor, human |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | EGFP is expressed in a small cluster of neurons in the hypothalamus. |
Strain of Origin | (B6.129P2-Hprtb-m3/J x 129S-Gt(ROSA)26Sortm1Sor/J)F1 |
Chromosome | X |
General Note | Germ line transmission of mutant cell line mEMS1774 has been established. |
Molecular Note | The Ple111-EGFP transgene (pEMS1417) was designed with the 544 bp Ple111 minipromoter (HCRT-A; derived from a subsection of the promoter from the human hypocretin (orexin) neuropeptide precursor (HCRT)) upstream of a minimal F5 mutant-frt site, an enhanced green fluorescent protein (with mutated TAA->TTA stop), a nuclear localization signal, a second minimal frt site, an SV40 early polyA signal, and a human HPRT complementary sequence (containing exon1, intron1, exon2, and part of intron2). This construct was targeted as a single copy knockin to the Hprtb-m3 mutant locus on the X chromosome. The promoter/regulatory regions of the human hypocretin (orexin) neuropeptide precursor (HCRT) directs expression of an enhanced green fluorescent protein (EGFP) to a small cluster of EGFP-positive neurons is found in the hypothalamus. EGFP-immunoreactivity fills the cytoplasm of these cells, highlighting the cell body as well as extensive dendritic processes. This label is very specific to the hypothalamus and not seen in any other cell populations in the brain. Double label immunocytochemistry shows that a majority of these EGFP-positive hypothalamic cells co-express with the HCRT (orexin) neuropeptide (the gene from which this MiniPromoter was designed). |
Mutations Made By | Elizabeth Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia |
The donating investigator recommends maintaining this strain by breeding heterozygous females with C57BL/6J inbred males.
When using the HCRT-EGFP mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #32927 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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