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| These piGAP transgenic reporter mice have expression of the eGFP-F-IRES-hPLAP dicistronic gene blocked by an upstream loxP-flanked STOP-polyA sequence. When bred to mice that express Cre recombinase, the resulting offspring will have the STOP-polyA sequence deleted in the cre-expressing tissue(s), permitting dicistronic expression of human Placental Alkaline Phosphatase (PLAP or ALPP) and farnesylated Enhanced Green Fluorescent Protein (eGFP-F; optimized to target expression to the cytoplasmic side of the plasma membrane). These piGAP transgenic reporter mice allow Cre-inducible, eGFP-F and hPLAP expression in multiple cell and tissue types. | |||||||||||
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Description
Strain Information
Type Coisogenic; Transgenic; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse Donating Investigator G. Giacomo Consalez, San Raffaele Scientific Institute (HSR) Description
Mice harboring the piGAP transgene are viable and fertile, with expression of the eGFP-F-IRES-hPLAP dicistronic gene blocked by an upstream loxP-flanked STOP-polyA sequence. When bred to mice that express Cre recombinase, the resulting offspring will have the STOP-polyA sequence deleted in the cre-expressing tissue(s), permitting dicistronic expression of eGFP-F-IRES-hPLAP. When transgene expression is induced in neurons, human Placental Alkaline Phosphatase (PLAP or ALPP) outlines axonal and dendritic projections and can be visualized by a simple histochemical reaction in fixed cells. Likewise, in vivo fluorescence of farnesylated Enhanced Green Fluorescent Protein (eGFP-F; optimized to target expression to the cytoplasmic side of the plasma membrane) labels axons, and dendrites throughout their length. Because both proteins localize alongside the neuronal surface, concomitant detection of cell body, neurites, and presynaptic and postsynaptic sites may be observed in the same neuron. These piGAP transgenic reporter mice allow Cre-inducible, eGFP-F and hPLAP expression in multiple cell and tissue types.For example, when bred to mice expressing Cre recombinase in neural tissues (such as Nestin-Cre (see Stock No. 003771) or CamKIIa-Cre (see Stock No. 005359)), the resulting offspring may be useful in studying axon guidance, circuit formation, and synaptic plasticity.
Development
The piGAP transgene was designed with a CMV enhancer/chicken beta-actin promoter (from pCAGGS vector) and loxP-flanked STOP-polyA sequence upstream of a farnesylated Enhanced Green Fluorescent Protein coding sequence (eGFP-F; from pEGFP-F plasmid (Clontech), eGFP fused to the carboxyl-terminal 20 amino acids of the human c-Ha-Ras protein, which targets GFP to the cytoplasmic side of the plasma membrane), internal ribosomal entry site (IRES), human Placental Alkaline Phosphatase cDNA sequence (PLAP or ALPP; from the pGTO plasmid), and SV40 polyA signal. This transgene was injected into the male pronucleus of fertilized oocytes of FVB/N females. Mice from the only founder line shown to express the transgene (founder line 2.6) were established and bred to FVB/N to establish the piGAP colony. Transgenic mice were then backcrossed to FVB/NCrl for at least 10 generations prior to arrival at The Jackson Laboratory. Upon arrival, transgenic mice may have been bred with FVB/NJ to establish the colony.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 001800 FVB/NJ | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (169 strains)
005703 B6.Cg-Tg(ACTFLPe)9205Dym/J 003800 B6;SJL-Tg(ACTFLPe)9205Dym/J 005145 C57BL/6-Tg(CAG-OVA)916Jen/J 005863 C57BL/6J-Tg(ACTB-DDAH1)1Jpck/J 002981 DBA/2-Tg(xstpx-lacZ)36And/J 003376 FVB/N-Tg(ACTB-cre)2Mrt/J 005438 STOCK Tg(CAG-Bgeo,-DsRed*MST)1Nagy/J
Additional Web Information
Cre-lox Systems
Fluorescent Proteins/lacZ Systems
Phenotype
Phenotype Information
assigned by genotype
Tg(CAG-EGFP,-ALPP)2.6Ggc/0
FVB/N-Tg(CAG-EGFP,-ALPP)2.6Ggc
- no phenotypic analysis
- *normal* no phenotypic analysis (MGI Ref ID J:136874)
This mouse can be used to support research in many areas including:
GFP relatedNeurobiology Research
Cre-lox System (loxP-flanked Sequences)
Cre-lox System (loxP-flanked Sequences: Test/Reporter)
Fluorescent protein expression in neural tissue
Research Tools
Cre-lox System (loxP-flanked Sequences: Test/Reporter)
Fluorescent Proteins
Genetics Research (Tissue/Cell Markers: Cre-lox System)
Genetics Research (Tissue/Cell Markers: multiple)
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(CAG-EGFP,-ALPP)2.6Ggc | ||
|---|---|---|---|
| Allele Name | transgene insertion 2.6, Giacomo Consalez | ||
| Allele Type | Transgenic (Reporter) | ||
| Common Name(s) | eGFP-F-IRES-hPLAP; piGAP; | ||
| Mutation Made By | G. Giacomo Consalez, San Raffaele Scientific Institute (HSR) | ||
| Strain of Origin | FVB/N | ||
| Site of Expression | GFP and alkaline phosphatase are expressed following cre-mediated excision of a transcriptional stop sequence. | ||
| Expressed Gene | ALPP, alkaline phosphatase, placental (Regan isozyme), human | ||
| 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 | ACTB, actin, beta, human | ||
| Molecular Note | The transgene was designed with a CMV enhancer/chicken beta-actin promoter (from pCAGGS vector) and loxP-flanked STOP-polyA sequence upstream of a farnesylated Enhanced Green Fluorescent Protein coding sequence (eGFP-F; from pEGFP-F plasmid (Clontech), eGFP fused to the carboxyl-terminal 20 amino acids of the human c-Ha-Ras protein, which targets GFP to the cytoplasmic side of the plasma membrane), internal ribosomal entry site (IRES), human Placental Alkaline Phosphatase cDNA sequence (PLAP or ALPP; from the pGTO plasmid), and SV40 polyA signal. [MGI Ref ID J:136874] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(CAG-EGFP,-ALPP)2.6Ggc, STD PCR, vers. 1
Helpful Links
Optimizing PCR Protocols
References
References
Selected Reference(s)
Badaloni A; Bonanomi D; Albieri I; Givogri I; Bongarzone E; Valtorta F; Consalez GG. 2007. Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis. Genesis 45(6):405-12. [PubMed: 17554764] [MGI Ref ID J:136874]
Health & husbandry
Health & Colony Maintenance Information
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, hemizygous mice may be bred to wildtype siblings or to FVB/NJ inbred mice. The donating investigator reports that attempts to generate homozygous mice were not successful and suggests that homozygosity may be lethal at some embryonic stage.
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
This strain is currently Under Development for Distribution Colony.
To register your interest in this strain go to the Strain Interest Form.
Estimated Available for Sale Date:
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.
View All Strains Under Development and On Hold
Supply Details
| Standard Supply | Under Development for Distribution Colony |
|---|---|
| Supply Notes |
|
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 001800 FVB/NJ | ||
| 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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