Description
The Lenti-X ProteoTuner Shield System N (w/ ZsGreen1) provides a unique method for quickly and directly regulating the amount of your protein of interest present in a cell, combined with versatile lentiviral delivery. The system utilizes a ligand-dependent destabilization domain (DD), derived from a 12 kDa mutant of the FKBP protein, and a membrane permeable stabilizing ligand, Shield1. Once cloned into the pLVX-PTuner Green Vector, the protein of interest is expressed with an N-terminal DD-tag and the fluorescent protein ZsGreen1. In the presence of Shield1, the DD-tagged protein is stabilized and will accumulate 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-tagged protein of interest is unstable. Consequently, removal of Shield1 (by splitting the cells into medium without Shield1) allows for destabilization and rapid degradation of the protein of interest. The extent of stabilization via Shield1 directly correlates with the amount of the ligand in the medium, so it is possible to fine-tune the amount of protein of interest present in the cell by controlling the amount of the Shield1 ligand.
