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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 Ple162-EGFP/cre mice, please see MMRRC #032922 at the MMRRC website. | |||||||||||||||||||
| These Ple162-EGFPCre;mEMS954 mice have the Ple162-EGFPCre 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 paired-like homeodomain 3 (PITX3) gene direct expression of an EGFPCre fusion protein to a discrete, non-dopaminergic cell population in the ventral midbrain that borders the fasciculus retroflexus and the dorsal edge of the main dopaminergic population of the ventral tegmental area (VTA), these Ple162-EGFPCre;mEMS954 mice may be useful in studying PITX3-expressing cells in the brain and diseases affecting the brain, including mesocorticolimbic dopamine system function in the drug and natural reward circuitry of the brain, cognition, motivation, drug addiction, and psychiatric disorders. | |||||||||||||||||||
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Description
Strain Information
Former Names B6.129P2(Cg)-Hprttm10(Ple162-EGFP/cre)Ems/Mmjax (Changed: 19-OCT-10 ) B6.129P2-Hprttm10(Ple162-EGFP/cre)Ems/J (Changed: 19-OCT-10 ) B6.129P2-Hprt1tm10(Ple162-EGFP/cre)Ems/J (Changed: 14-DEC-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 Heterozygote x Hemizygote (Female x Male) 13-JAN-10 Species laboratory mouse Generation N6+N2F3pN1
Generation DefinitionsDonating Investigator Elizabeth Simpson, University of British Columbia Description
These Ple162-EGFPCre;mEMS954 mice have the Ple162-EGFPCre 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 paired-like homeodomain 3 (PITX3) gene directing expression of an enhanced green fluorescent protein/Cre recombinase fusion protein (EGFPCre). These Ple162-EGFPCre;mEMS954 mice may be useful in studying PITX3-expressing cells in the brain and diseases affecting the brain, including mesocorticolimbic dopamine system function in the drug and natural reward circuitry of the brain, cognition, motivation, drug addiction, and psychiatric disorders.For Ple162-EGFPCre;mEMS954 expression information, see The Pleiades Promoter Project website (Ple162 Promoter (pEMS1148)).
The donating investigator reports:
When these Ple162-EGFPCre;mEMS954 mice are bred to also harbor the Gt(ROSA)26Sortm1Sor mutant allele, strong Cre recombinase activity (βGal staining) is detected in a discrete neuronal-like cell population in the ventral midbrain. In the mature brain, βGal-positive cells are found surrounding the fasciculus retroflexus. Double labeling for ?Gal and tyrosine hydroxylase shows that the βGal-positive cells are not dopaminergic cells, but do border the dorsal edge of the main dopaminergic population of the VTA. A developmental analysis showed clear βGal labeling of ventral midbrain cells in embryonic day 11.5 (E11.5) and E15.5 embryos as well as postnatal day 0 (P) and P7 pups, in the regions from and through which the dopaminergic population arises and migrates, respectively. Active expression of the EGFP/cre reporter gene product, however, cannot be detected in these cells by EGFP immunostaining, indicating βGal expression is an historical marker of expression of the EGFP/cre reporter. Additionally, a substantial non-CNS staining was detected in muscle tissue of the abdominal wall, limb buds and posterior paravertebral tissues in E11.5 whole embryos as expected for the PITX3 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 Ple162-EGFPCre transgene (pEMS1148) was designed with the 3636 bp Ple162 minipromoter (PITX3-D; derived from an intronic candidate regulatory region and a segment of the human paired-like homeodomain 3 (PITX3) promoter) upstream of a minimal F5 mutant-frt site, an EGFP/Cre fusion protein (enhanced green fluorescent protein (with mutated stop) and Cre recombinase), 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 mEMS21 embryonic stem cells (derived from 129P2/OlaHsd-derived E14TG2a ES cells; a karyotypically male ES cell line harboring the Hprtb-m3 mutation on the X chromosome). Correctly targeted embryonic stem cells were microinjected into recipient mice cells. The resulting chimeric mice (founder line mEMS954) were bred to B6.129S4-Gt(ROSA)26Sortm1Sor/J mice (Stock No. 003474) to establish the mutant colony. These Ple162-EGFPCre;mEMS954 mutant mice were bred with B6.129S4-Gt(ROSA)26Sortm1Sor/J mice (and/or C57BL/6J inbred mice (see Stock No. 000664)) for at least five generations prior to arrival at The Jackson Laboratory. Upon arrival, Ple162-EGFPCre;mEMS954 mutant mice were backcrossed to C57BL/6J inbred mice (selecting away from the Gt(ROSA)26Sortm1Sor mutation) to establish this congenic strain.
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
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Additional Web Information
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Phenotype
Phenotype Information
assigned by genotype
Hprttm10(Ple162-EGFP/cre)Ems/Y
B6.129P2(129S4)-Hprttm10(Ple162-EGFP/cre)Ems
- 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
Cre-lox System
Cre Recombinase expression in neural tissue
Research Tools
Cre-lox System
Cre Recombinase Expression
Developmental Biology Research
Cre-lox System
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: neurons
Neurobiology Research
cell marker
cre relatedResearch Tools
Fluorescent Proteins
Cre-lox System
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Hprttm10(Ple162-EGFP/cre)Ems | ||
|---|---|---|---|
| Allele Name | targeted mutation 10, Elizabeth M Simpson | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | Hprt1tm10(Ple162-EGFPcre;mEMS954)Ems; Hprt1tm10(mEMS954)Ems; PITX3-EGFP/cre; | ||
| Mutation Made By | Elizabeth Simpson, University of British Columbia | ||
| Strain of Origin | 129P2/OlaHsd | ||
| ES Cell Line Name | E14TG2a | ||
| ES Cell Line Strain | 129P2/OlaHsd | ||
| 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. | |||
| Expressed Gene | cre, cre recombinase, bacteriophage P1 | ||
| Cre recombinase is an enzyme derived from the bacteriophage P1 that specifically recognizes loxP sites. Cre has been shown to effectively mediate the excision of DNA located between loxP sites. After the excision event, the DNA ends recombine leaving a single loxP site in place of the intervening sequence. | |||
| Promoter | PITX3, paired-like homeodomain 3, human | ||
| Associated Marker Note | Expressed-count: 2Ex1-Source: JellyfishEx1-Gene: EGFPEx2-Source: Bacteriophage P1Ex2-Gene: creDriver-count: 1Dr1-Source: HumanDr1-Gene: 5309 | ||
| Driver Note | PITX3 | ||
| General Note | Germ line transmission of mutant cell line mEMS954 has been established. | ||
| Molecular Note | The Ple162-EGFPCre transgene (pEMS1148) was designed with the 3636 bp Ple162 minipromoter (PITX3-D; derived from an intronic candidate regulatory region and a segment of the human paired-like homeodomain 3 (PITX3) promoter) upstream of a minimal F5 mutant-frt site, an EGFP/Cre fusion protein (enhanced green fluorescent protein (with mutated stop) and Cre recombinase), 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). [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
Hprt1tm10(Ple162-EGFP/cre)Ems STD PCR, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
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]
Additional References
Hprttm10(Ple162-EGFP/cre)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]
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). Mating System Heterozygote x Hemizygote (Female x Male) 13-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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