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| 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 1 (Hprt1) 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
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 See Colony Maintenance Species laboratory mouse Generation N5+N1F (22-JUL-09) Donating 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 1 (Hprt1) 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 Hprt1b-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 Hprt1b-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 | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (225 strains)
009114 B6.129P2(129S4)-Hprt1tm14(Ple103-EGFP/cre)Ems/J 008877 B6.129P2-Hprt1tm12(Ple177-EGFP/cre)Ems/J 008709 B6.129P2-Hprt1tm9(Ple178-EGFP/cre)Ems/J 008876 B6;129-Hprt1tm11(Ple176-EGFP/cre)Ems/J
004302 129S1-Hprt1tm1(cre)Mnn/J 002027 129S8/SvEv-Gpi1c Hprt1b-m2/J 002171 B6.129P2-Hprt1b-m3/J 008877 B6.129P2-Hprt1tm12(Ple177-EGFP/cre)Ems/J 008706 B6.129P2-Hprt1tm4(Ple88-EGFP)Ems/J 008707 B6.129P2-Hprt1tm7(Ple185-EGFP)Ems/J 008708 B6.129P2-Hprt1tm8(Ple151-EGFP)Ems/J 008709 B6.129P2-Hprt1tm9(Ple178-EGFP/cre)Ems/J 008876 B6;129-Hprt1tm11(Ple176-EGFP/cre)Ems/J 003138 B6;129-Hprt1tm1Detl/J 000807 RBJ/DnJ
Additional Web Information
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Phenotype
Phenotype Information
This mouse can be used to support research in many areas including:
GFP relatedNeurobiology Research
Cre-lox System
Cre Recombinase expression in neural tissue
Fluorescent protein expression in neural tissue
Research Tools
Cre-lox System
Cre Recombinase Expression
Developmental Biology Research
Cre-lox System
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
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: Cre-lox System
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Hprt1tm10(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; | ||
| Strain of Origin | 129P2/OlaHsd | ||
| ES Cell Line Name | mEMS21 | ||
| ES Cell Line Strain | 129P2/OlaHsd | ||
| 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. | |||
| 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 | PITX3, paired-like homeodomain 3, human | ||
| Driver Note | Ple162 | ||
| 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). | ||
| Gene Symbol and Name | Hprt1, hypoxanthine guanine phosphoribosyl transferase 1 | ||
| Chromosome | X | ||
| Gene Common Name(s) | C81579; HGPRT; HPGRT; HPRT; Hgprtase; MGC112554; expressed sequence C81579; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Hprt1tm10(Ple162-EGFP/cre)Ems STD PCR, Standard PCR
Gt(ROSA)26Sortm1sor STD, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
The Pleiades Promoter Project, URL: http://pleiades.org/index.php. 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 donating investigator recommends maintaining this strain by breeding heterozygous females with C57BL/6J inbred males (Stock No. 000664). Mating System See above Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
This strain is currently Under Development for Production.
To register your interest in this strain go to the Strain Interest Form.
Estimated Available for Sale Date: 23-NOV-09
Please note: Estimated available for sale dates are provided to keep customers better informed on strains under development. Please note that our Colony Managers routinely monitor the target date and edit it based on breeding performance and other factors. The length of time it takes to make a new strain available for sale depends on genotype, age, number of animals sent by the Donating Investigator, breeding performance, additional strain development (backcrossing, making homozygous), and anticipated demand for the strain/interest registered.
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Supply Details
| Standard Supply | Under Development for Distribution Colony |
|---|---|
| Supply Notes |
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Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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