Takara ProteoTuner Shield System C \Each

The ProteoTuner Shield System C utilizes a ligand-dependent destabilization domain (DD-C) and a membrane-permeant stabilizing ligand, Shield1, to quickly and directly regulate the amount of your protein of interest present in a cell or organism. First, your gene of interest is cloned into the pPTunerC Vector and expressed as a fusion with the destabilization domain (DD-C). Then the Shield1 ligand is either added or withheld, depending on your experiment.

In the presence of Shield1, the DD-C-tagged protein is stabilized and accumulates inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15–30 minutes after the addition of Shield1. In the absence of Shield1, the DD-C-tagged protein of interest is unstable. Consequently, removal of Shield1 allows for active and controllable degradation of the protein of interest. The extent of stabilization via Shield1 can be adjusted, and directly correlates with the amount of the ligand in the medium. Therefore, you can tune the level of your protein of interest present in the cell, by controlling the amount of the Shield1 ligand.

Ligand-dependent, targeted and reversible protein stabilization.A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present.

Control of actin stability with the ProteoTuner system.In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).