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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 Ple88-EGFP mice, please see MMRRC #032918 at the MMRRC website. | ||||||||||||||||||||||
| These Ple88-EGFP;mEMS1671 mice have the Ple88-EGFP transgene targeted as a single copy "knockin" into the upstream region of hypoxanthine guanine phosphoribosyl transferase (Hprt) locus on the X chromosome. As the promoter/regulatory regions of the human glial fibrillary acidic protein (GFAP) gene direct expression of enhanced green fluorescent protein (EGFP), these Ple88-EGFP;mEMS1671 mice may be useful in studying GFAP-expressing cells in the brain and diseases affecting the brain. | ||||||||||||||||||||||
- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
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Description
Strain Information
Former Names B6.129P2-Hprttm4(Ple88-EGFP)Ems/J (Changed: 19-OCT-10 ) B6.129P2-Hprt1tm4(Ple88-EGFP)Ems/J (Changed: 14-DEC-09 ) B6.129P2-Hprt1tm4(GFAP-EGFP)Ems/J (Changed: 10-MAR-09 ) Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System Homozygote x Hemizygote (Female x Male) 08-JAN-10 Species laboratory mouse Generation N4+N1F7 (26-OCT-09)
Generation DefinitionsDonating Investigator Elizabeth Simpson, University of British Columbia Description
These Ple88-EGFP;mEMS1671 mice have the Ple88-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 glial fibrillary acidic protein (GFAP) gene directing expression of enhanced green fluorescent protein (EGFP). These Ple88-EGFP;mEMS1671 mice may be useful in studying GFAP-expressing cells in the brain and diseases affecting the brain.For Ple88-EGFP;mEMS1671 expression information, see The Pleiades Promoter Project website (Ple88 Promoter (pEMS1375)).
The donating investigator reports:
EGFP-positive cells are present throughout the brain of Ple88-EGFP mice, presenting with varying staining characteristics. In most cells, EGFP immunoreactivity fills the cell bodies and processes, revealing their full morphology and indicating the glial nature of these labelled cells. As expected in the white matter, there are astroglial-like profiles found in the corpus callosum, alveus, fimbria, cerebellar white matter and other fibre tracts in the brain. In the neuropil of the grey matter throughout the brain, there are EGFP-positive cells that have an immunopositivity meshwork that surrounds each cell, characteristic of the processes of a protoplasmic astrocyte. Specialized glial cell types that form the glia limitans and the Bergman glial fibers also express the EGFP reporter. In the eye, numerous cells are observed in the optic nerve and the proximal segment as the nerve enters the retina much like typical astrocytic staining in this region. There is not a complete overlap between cells in the brain that co-express the GFAP target gene and the EGFP reporter gene. Thus, expression from the Ple88 promoter reflects an expression pattern beyond that of the endogenous GFAP gene.These mice were created and deposited by The Pleiades Promoter Project (Centre for Molecular Medicine and Therapeutics, University of British Columbia); their goal is to generate 160 fully characterized, human DNA promoters of less than 4 kb (MiniPromoters) to drive gene expression in defined brain regions of therapeutic interest for studying disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease, Amyotrophic Lateral Sclerosis (Lou Gehrig's disease), Multiple Sclerosis, Spinocerebellar Ataxia, Depression, Autism, and Cancer.
Development
The Ple88-EGFP transgene (pEMS1375) was designed with a Ple88 minipromoter (GFAP-A; a 2210 bp promoter region of the human glial fibrillary acidic protein (GFAP) gene) 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 mEMS1671) were bred to B6(Cg)-Tyrc-2J/J females (B6-albino; Stock No. 000058). Female progeny (obligate HprtPle88-EGFP;mEMS1671 heterozygotes) also harboring a Tyr+ allele in combination with the Aw (agouti, white belly) and a (non-agouti/black) coat color alleles were identified and bred with C57BL/6J inbred males (Stock No. 000664). Next, hemizygous HprtPle88-EGFP;mEMS1671 mice were bred with C57BL/6J inbred mice for at least three generations prior to arrival at The Jackson Laboratory. Upon arrival, these mice were bred with C57BL/6J inbred mice for at least one generation to establish the live colony.For more information, see The Pleiades Promoter Project website (Ple88 Promoter (pEMS1375)).
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
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Additional Web Information
Fluorescent Proteins/lacZ Systems
JAX® NOTES, Spring 2009; 513. Pleiades Promoter Project mice shine at The Jackson Laboratory.
Phenotype
Phenotype Information
assigned by genotype
Hprttm4(Ple88-EGFP)Ems/Y
B6.129P2(Cg)-Hprttm4(Ple88-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: astrocytes
Tissue/Cell Markers: astrocytes, neurons
Tissue/Cell Markers: glial cells
Tissue/Cell Markers: neurons
Neurobiology Research
cell marker
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Hprttm4(Ple88-EGFP)Ems | ||
|---|---|---|---|
| Allele Name | targeted mutation 4, Elizabeth M Simpson | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | Hprt1tm4(GFAP-EGFP)Ems; Hprt1tm4(Ple88-EGFP;mEMS1671)Ems; Hprt1tm4(mEMS1671)Ems; Ple88;GFAP-A;mEMS1671; | ||
| Mutation Made By | Elizabeth Simpson, University of British Columbia | ||
| Strain of Origin | (B6.129P2-Hprt | ||
| ES Cell Line Name | mEMS1204 | ||
| ES Cell Line Strain | (B6.129P2-Hprt | ||
| Site of Expression | neural white matter glia and astrocytes | ||
| 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. | |||
| Promoter | GFAP, glial fibrillary acidic protein, human | ||
| Associated Marker Note | Expressed-count: 1Ex1-Source: JellyfishEx1-Gene: EGFPDriver-count: 1Dr1-Source: HumanDr1-Gene: 2670 | ||
| General Note | Germ line transmission of mutant cell line mEMS1671 has been established. | ||
| Molecular Note |
The Ple88-EGFP transgene (pEMS1375) was designed with a Ple88 minipromoter (GFAP-A; a 2210 bp promoter region of the human glial fibrillary acidic protein (GFAP) gene) 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. In most cells, EGFP immunoreactivity fills the cell bodies and processes; revealing their full morphology and indicating the glial nature of the cell types. There is EGFP expression inA): astroglial-like cells in the forebrain and cortex, hippocampus and alvius, the cerebellar white matter, as well as the midbrain, brainstem, and eye, B): nascent neuronal/glial progenitor cells in the developing forebrain (including rostral cortical subventricular zone and the rostral migratory stream, and C): numerous cells in the optic nerve and the proximal segment as the nerve enters the retina (much like typical astrocytic staining). While EGFP and GFAP co-expression is observed, not all EGFP+ cells are GFAP+; reflecting the possibility that the Ple88 minipromoter may express in non-glial cell types. [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
Hprt1tm4(Ple88-EGFP)Ems STD PCR, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
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]
Additional References
Hprttm4(Ple88-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]
Health & husbandry
Health & Colony Maintenance Information
Colony Maintenance
Breeding & Husbandry The mutant allele is located on the X chromosome. When maintaining a live colony, homozygous females may be bred with hemizygous males. Mating System Homozygote x Hemizygote (Female x Male) 08-JAN-10
Purchasing information
Pricing, Supply Level & Notes, Controls
This strain is currently Transferred.
General Supply Notes
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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