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).
Nico Dantuma, Karolinska Institutet
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter) |
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).
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.
Expressed Gene | GFP, Green Fluorescent Protein, |
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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. |
Allele Name | transgene insertion 2, Nico Dantuma |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | Tg(ACTB-Ub*G76V/GFP)2Dant; Tg(CAG-Ub*G76V/GFP)2Dant; UbG76V-GFP/2; UbG76V-GFP/2; UPS reporter |
Gene Symbol and Name | Tg(CAG-Ub*G76V/GFP)2Dant, transgene insertion 2, Nico Dantuma |
Gene Synonym(s) | |
Promoter | ACTB, actin, beta, chicken |
Expressed Gene | GFP, Green Fluorescent Protein, |
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. |
Strain of Origin | (CBA x C57BL/6)F1 |
Chromosome | UN |
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. |
Mutations Made By | Nico Dantuma, Karolinska Institutet |
When maintaining a live colony, hemizygotes may be bred to wildtype siblings or to C57BL/6NJ (Stock No. 005304) inbred mice.
When using the B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #008112 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous or Non carrier for Tg(CAG-Ub*G76V/GFP)2Dant |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J Frozen Embryo | $3373.50 |
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