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Protein Degradation and Control Technology

How it works

Gene regulation studies are essential to monitor the effect of the presence and absence of a specific protein of interest. Reversible control of a specific protein expression is a powerful tool for evaluating protein function. Small molecule dependent (Shield ligands) Destabilization Domain (DD) system allows to post translationally regulate the amount of protein of interest present in a cell. It utilizes a modified protein domain that is co-dependent on a membrane permeable stabilizing ligand, Shield1. The DD system is based on a 12 kDa mutant of the FKBP protein, which is expressed as a tag onto the N-terminus of your protein of interest cloned into the vector. In the presence of Shield1, the DD-tagged protein is stabilized and starts accumulating 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. However, in the absence of the protective ligand switch Shield1, the DD-tagged protein of interest is rapidly degraded. The magnitude of stabilization can be controlled via the concentration of Shield1 in the medium.

“A Rapid, Reversible and Tunable Method to Regulate Protein Function in Living Cells Using Synthetic Small Molecule.” Banaszynski, L. A.; Chen, L.-C.; Maynard-Smith, L.A.; Ooi, A. G. L.; Wandless, T.J. Cell, 2006, 126, 995-1004.

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