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| Mouse strains created by the Pleiades Promoter Project (Dr. Elizabeth M. Simpson, Centre for Molecular Medicine and Therapeutics, University of British Columbia, Canada) are now available through the Mutant Mouse Regional Resource Center (MMRRC) at The Jackson Laboratory. For the Ple111-EGFP mice, please see MMRRC #032927 at the MMRRC website. | ||||||||||||||||||||||
| 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. | ||||||||||||||||||||||
- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
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Description
Strain Information
Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Species laboratory mouse Generation N4pN1
Generation DefinitionsDonating Investigator Elizabeth Simpson, University of British Columbia Description
These Ple111-EGFP;mEMS1774 mice have the Ple111-EGFP transgene targeted as a single copy "knockin" into the upstream region of hypoxanthine guanine phosphoribosyl transferase (Hprt) locus on the X chromosome. Heterozygous females and hemizygous males are viable and fertile, with the promoter/regulatory regions of the human hypocretin (orexin) neuropeptide precursor (HCRT) directing expression of an enhanced green fluorescent protein (EGFP). These Ple111-EGFP;mEMS1774 mice may be useful in studying HCRT -expressing cells in the brain and diseases affecting the brain.For Ple111-EGFP;mEMS1774 expression information, see The Pleiades Promoter Project website (Ple111 Promoter (pEMS1417)).
The donating investigator reports:
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).These mice were created and deposited by The Pleiades Promoter Project (Centre for Molecular Medicine and Therapeutics, University of British Columbia).
Development
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 via electroporation into mEMS1204 embryonic stem cells (generated from F1 generation male pups born of C57BL/6-congenic females homozygous for a Hprtb-m3 mutation [originally derived from 129P2/OlaHsd-derived E14TG2a embryonic stem cells] X 129S-Gt(ROSA)26Sortm1Sor heterozygous males (see Stock No. 002171 and 003310, respectively)). Correctly targeted embryonic stem cells were microinjected into recipient mice cells. The resulting chimeric males (founder line mEMS1774) were bred to B6(Cg)-Tyrc-2J/J females (B6-albino; Stock No. 000058) to establish the mutant colony. The donating investigator reports that these Ple111-EGFP;mEMS1774 mutant mice were bred with C57BL/6J wildtype mice for four generations (and selectively removed the Gt(ROSA)26Sortm1Sor mutation) prior to sending to The Jackson Laboratory. Upon arrival, Ple111-EGFP;mEMS1774 mutant mice were used to cryopreserve sperm. When rederiving a live colony, sperm will be used along with oocytes from C57BL/6J inbred mice (Stock No. 000664) to generate obligate heterozygous females, and these females will be bred at least one generation to C57BL/6J inbred mice to establish the colony.For more information, see The Pleiades Promoter Project website (Ple111 Promoter (pEMS1417)).
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
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Additional Web Information
Fluorescent Proteins/lacZ Systems
Phenotype
Phenotype Information
assigned by genotype
Hprttm15(Ple111-EGFP)Ems/Y
B6.129P2(Cg)-Hprttm15(Ple111-EGFP)Ems/Mmjax
- normal phenotype
- no abnormal phenotype detected
- heterozygous females and hemizygous males are viable and fertile (MGI Ref ID J:145689)
This mouse can be used to support research in many areas including:
GFP relatedNeurobiology Research
Fluorescent protein expression in neural tissue
Research Tools
Fluorescent Proteins
Genetics Research
Tissue/Cell Markers
Tissue/Cell Markers: glial cells
Tissue/Cell Markers: glial cells, oligodendrocytes, Schwann cells
Tissue/Cell Markers: neurons
Neurobiology Research
cell marker
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Hprttm15(Ple111-EGFP)Ems | ||
|---|---|---|---|
| Allele Name | targeted mutation 15, Elizabeth M Simpson | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | HCRT-EGFP; Hprt1tm15(Ple111-EGFP;mEMS1774)Ems; Hprt1tm15(mEMS1774)Ems; Ple111; | ||
| Mutation Made By | Elizabeth Simpson, University of British Columbia | ||
| Strain of Origin | (B6.129P2-Hprt | ||
| Expressed Gene | GFP, Green Fluorescent Protein, jellyfish | ||
| Green Fluorescent Protein (GFP), derived from the jellyfish Aequorea victoria, is a versatile reporter molecule which has found use in many biological applications. In some constructs the original molecule has been modified in order to enhance its fluorescence intensity (EGFP, enhanced GFP). When utilized in a transgenic construct, tissue expressing sufficient amounts of GFP will fluoresce when exposed to a 488 nm light source. | |||
| Associated Marker Note | Expressed-count: 1Ex1-Source: JellyfishEx1-Gene: EGFPDriver-count: 1Dr1-Source: HumanDr1-Gene: 3060 | ||
| 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). [MGI Ref ID J:144244] [MGI Ref ID J:164356] | ||
| Gene Symbol and Name | Hprt, hypoxanthine guanine phosphoribosyl transferase | ||
| Chromosome | X | ||
| Gene Common Name(s) | C81579; HGPRT; Hgprtase; Hprt1; expressed sequence C81579; hypoxanthine guanine phosphoribosyl transferase 1; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Gt(ROSA)26Sortm1sor STD, Robotic STD
Gt(ROSA)26Sortm1sor STD, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Additional References
Hprttm15(Ple111-EGFP)Ems relatedPortales-Casamar E; Swanson DJ; Liu L; de Leeuw CN; Banks KG; Ho Sui SJ; Fulton DL; Ali J; Amirabbasi M; Arenillas DJ; Babyak N; Black SF; Bonaguro RJ; Brauer E; Candido TR; Castellarin M; Chen J; Chen Y; Cheng JC; Chopra V; Docking TR; Dreolini L; D'Souza CA; Flynn EK; Glenn R; Hatakka K; Hearty TG; Imanian B; Jiang S; Khorasan-zadeh S; Komljenovic I; Laprise S; Liao NY; Lim JS; Lithwick S; Liu F; Liu J; Lu M; McConechy M; McLeod AJ; Milisavljevic M; Mis J; O'Connor K; Palma B; Palmquist DL; Simpson EM:. 2010. A regulatory toolbox of MiniPromoters to drive selective expression in the brain. Proc Natl Acad Sci U S A 107(38):16589-94. [PubMed: 20807748] [MGI Ref ID J:164356]
The Pleiades Promoter Project. 2009. Generation of reporter and cre-expressing targeted transgenic mice in the Hprt1 gene using human MiniPromoters that drive region- and cell-specific gene expression in the mouse brain MGI Direct Data Submission :. [MGI Ref ID J:145689]
Yang GS; Banks KG; Bonaguro RJ; Wilson G; Dreolini L; de Leeuw CN; Liu L; Swanson DJ; Goldowitz D; Holt RA; Simpson EM. 2009. Next generation tools for high-throughput promoter and expression analysis employing single-copy knock-ins at the Hprt1 locus. Genomics 93(3):196-204. [PubMed: 18950699] [MGI Ref ID J:144244]
Health & husbandry
Health & Colony Maintenance Information
Colony Maintenance
Breeding & Husbandry The donating investigator recommends maintaining this strain by breeding heterozygous females with C57BL/6J inbred males (Stock No. 000664).
Purchasing information
Pricing, Supply Level & Notes, Controls
This strain is currently Transferred.
General Supply Notes
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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