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| Both ubiquitin/proteasome system reporter (UPS reporter) founder lines, UbG76V-GFP/1 (Stock No. 008111) and UbG76V-GFP/2 (Stock No. 008112), may be useful for monitoring the role of ubiquitin/proteasome-dependent proteolysis in diverse disorders, and for efficacy trials testing the effect of compounds on the ubiquitin/proteasome system (including the effect of proteasome inhibitors as novel anticancer drugs). | |||||||||||||||
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Description
Strain Information
Former Names B6.Cg-Tg(ACTB-Ub*G76V/GFP)2Dant/J (Changed: 08-MAY-08 ) Type Congenic; Mutant Strain; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System Noncarrier x Hemizygote (Female x Male) 22-JUL-08 Species laboratory mouse Generation N31+F2 (06-JAN-09) Donating Investigator Nico Dantuma, Karolinska Institutet Description
Hemizygous transgenic mice are viable and fertile; they contain the green fluorescent protein (GFP) fused to a constitutively active degradation signal (UbG76V). Transgenic transcripts are detected in all tissues examined; however no GFP protein expression is detected due to the G76V substitution which leads to its ubiquitination and proteasomal degradation. Following administration of proteasome inhibitors, UbG76V accumulates and GFP-derived fluorescence is readily apparent, evidence of an impaired ubiquitin/proteasome system. This strain and UbG76V-GFP/1 (Stock No. 008111) have similar expression patterns, but this line (UbG76V-GFP/2) shows lower transgene expression and is not reported to display background fluorescence. These strains may be useful for monitoring ubiquitin/proteasome-dependent proteolysis in diverse disorders, and in efficacy trials for monitoring the effect of compounds on the ubiquitin/proteasome system (for example, in tests with proteasome inhibitors as novel anticancer drugs).Development
The UbG76V-GFP transgene was designed with a chicken beta-actin promoter (and cytomegalovirus (CMV) immediate early enhancer) upstream of a green fluorescent protein (GFP) fused to a mutant ubiquitin moiety (UbG76V), a constitutively active proteasome degradation signal. The transgene was microinjected into the pronucleus of fertilized (CBAxC57BL/6)F1 oocytes, and the transgenic offspring e bred to C57BL/6N. Founder line 2 (UbG76V-GFP/2) mice, with lower expression of the transgene than line 1, were backcrossed to C57BL/6N for approximately 30 generations prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred with C57BL/6NJ (Stock No. 005304) to establish the colony.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 005304 C57BL/6NJ | ||
| Considerations for Choosing Controls | ||
Related Strains
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Additional Web Information
Fluorescent Proteins/lacZ Systems
Phenotype
Phenotype Information
assigned by genotype
Tg(CAG-Ub*G76V/GFP)2Dant/0
B6N.Cg-Tg(CAG-Ub*G76V/GFP)2Dant
- normal phenotype
- no abnormal phenotype detected (MGI Ref ID J:127085)
- hemizygous mice are viable and fertile
This mouse can be used to support research in many areas including:
GFP relatedCancer Research
Other
Cell Biology Research
Post-translational Processing
Protein Processing
degradation
Neurobiology Research
Fluorescent protein expression in neural tissue
Research Tools
Cancer Research
Cell Biology Research
Fluorescent Proteins
Toxicology Research
drug/compound testing
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(CAG-Ub*G76V/GFP)2Dant | ||
|---|---|---|---|
| Allele Name | transgene insertion 2, Nico Dantuma | ||
| Allele Type | Transgenic (Reporter) | ||
| Common Name(s) | Tg(ACTB-Ub*G76V/GFP)2Dant; Tg(CAG-Ub*G76V/GFP)2Dant; UPS reporter; UbG76V-GFP/2; UbG76V-GFP/2; | ||
| Mutation Made By | Nico Dantuma, Karolinska Institutet | ||
| Strain of Origin | (CBA x C57BL/6)F1 | ||
| Site of Expression | Transgene transcripts are detected in all tissues examined; however the G76V substitution leads to degradation, of the GFP. Following administration of proteasome inhibitors, UbG76V accumulates and GFP-derived fluorescence is apparent. This line shows lower transgene expression and is not reported to display background fluorescence. | ||
| 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, chicken | ||
| Molecular Note | The UbG76V-GFP transgene was designed with a chicken beta-actin promoter (and cytomegalovirus (CMV) immediate early enhancer) upstream of a green fluorescent protein (GFP) fused to a mutant ubiquitin moiety (UbG76V). This transgene was microinjected into the pronucleus of fertilized (CBA x C57BL/6)F1 oocytes. Although transcripts of the UbG76V-GFP fusion gene are detected in all tissues examined, the G76V substitution prevents removal of the ubiquitin moiety leading to efficient ubiquitination and proteasomal degradation of the fusion protein (and thus no fluorescence) in these tissues. Line 2 with low expression was maintained by backcross to C57BL/6N mice. [MGI Ref ID J:127085] [MGI Ref ID J:127087] | ||
| Gene Symbol and Name | Tg(CAG-Ub*G76V/GFP)2Dant, transgene insertion 2, Nico Dantuma | ||
| Chromosome | UN | ||
| Gene Common Name(s) | Tg(ACTB-Ub*G76V/GFP)2Dant; UPS reporter; UbG76V-GFP/2; UbG76V-GFP/2; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(CAG-Ub*G76V/GFP)1Dant, Tg(CAG-Ub*G76V/GFP)2Dant, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Bowman AB; Yoo SY; Dantuma NP; Zoghbi HY. 2005. Neuronal dysfunction in a polyglutamine disease model occurs in the absence of ubiquitin-proteasome system impairment and inversely correlates with the degree of nuclear inclusion formation. Hum Mol Genet 14(5):679-91. [PubMed: 15661755] [MGI Ref ID J:96393]
Lindsten K; Menendez-Benito V; Masucci MG; Dantuma NP. 2003. A transgenic mouse model of the ubiquitin/proteasome system. Nat Biotechnol 21(8):897-902. [PubMed: 12872133] [MGI Ref ID J:127085]
Additional References
Tg(CAG-Ub*G76V/GFP)2Dant relatedCheroni C; Marino M; Tortarolo M; Veglianese P; De Biasi S; Fontana E; Zuccarello LV; Maynard CJ; Dantuma NP; Bendotti C. 2009. Functional alterations of the ubiquitin-proteasome system in motor neurons of a mouse model of familial amyotrophic lateral sclerosis. Hum Mol Genet 18(1):82-96. [PubMed: 18826962] [MGI Ref ID J:142811]
Dantuma NP; Lindsten K; Glas R; Jellne M; Masucci MG. 2000. Short-lived green fluorescent proteins for quantifying ubiquitin/proteasome-dependent proteolysis in living cells. Nat Biotechnol 18(5):538-43. [PubMed: 10802622] [MGI Ref ID J:127087]
Lindsten K; de Vrij FM; Verhoef LG; Fischer DF; van Leeuwen FW; Hol EM; Masucci MG; Dantuma NP. 2002. Mutant ubiquitin found in neurodegenerative disorders is a ubiquitin fusion degradation substrate that blocks proteasomal degradation. J Cell Biol 157(3):417-27. [PubMed: 11980917] [MGI Ref ID J:127086]
Mammucari C; Milan G; Romanello V; Masiero E; Rudolf R; Del Piccolo P; Burden SJ; Di Lisi R; Sandri C; Zhao J; Goldberg AL; Schiaffino S; Sandri M. 2007. FoxO3 controls autophagy in skeletal muscle in vivo. Cell Metab 6(6):458-71. [PubMed: 18054315] [MGI Ref ID J:130442]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX12
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, hemizygotes may be bred to wildtype siblings or to C57BL/6NJ (Stock No. 005304) inbred mice. Mating System Noncarrier x Hemizygote (Female x Male) 22-JUL-08 Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
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Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $243.50 Female or Male Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant
Pairs /Price (US dollars $) Pair Genotype $297.85 Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant x Noncarrier $297.85 Noncarrier x Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant
Additional Supply Details
| Pricing for International shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $316.60 Female or Male Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant
Pairs /Price (US dollars $) Pair Genotype $387.30 Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant x Noncarrier $387.30 Noncarrier x Hemizygous for Tg(CAG-Ub*G76V/GFP)2Dant
Additional Supply Details
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Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of approximately nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within two business days following order placement. |
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| Supply Notes |
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Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 005304 C57BL/6NJ | ||
| 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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