B6.Cg-Tg(CAG-Ub*G76V/GFP)2Dant/J

Stock number: 008112
Research areas: Cancer Research, Cell Biology Research, Neurobiology Research, Research Tools

Description

Both ubiquitin/proteasome system reporter (UPS reporter) founder lines, Ub^G76V-GFP/1 (Stock No. 008111) and Ub^G76V-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).

Detailed description

Hemizygous transgenic mice are viable and fertile; they contain the green fluorescent protein (GFP) fused to a constitutively active degradation signal (Ub^G76V). 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, Ub^G76V accumulates and GFP-derived fluorescence is readily apparent, evidence of an impaired ubiquitin/proteasome system. This strain and Ub^G76V-GFP/1 (Stock No. 008111) have similar expression patterns, but this line (Ub^G76V-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 Ub^G76V-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 (Ub^G76V), 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 (Ub^G76V-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.

Allele

Symbol: Tg(CAG-Ub*G76V/GFP)2Dant
Name: transgene insertion 2, Nico Dantuma
MGI accession ID: MGI:3762796
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(CAG-Ub*G76V/GFP)2Dant
Marker name: transgene insertion 2, Nico Dantuma
Chromosome: UN
Strain of origin: (CBA x C57BL/6)F1
Expressed genes: 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, Ub^G76V accumulates and GFP-derived fluorescence is apparent. This line shows lower transgene expression and is not reported to display background fluorescence.
Molecular note: The Ub^G76V-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 Ub^G76V-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.