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Protein ubiquitination is a dynamic process, depending on a tightly regulated balance between the activity of ubiquitin ligases and their antagonists, the ubiquitin-specific proteases or deubiquitinating enzymes. The family of ubiquitin ligases has been studied intensively and it is well established that their deregulation contributes to diverse disease processes, including cancer. Much less is known about the function and regulation of the large group of deubiquitinating enzymes. This chapter describes how RNA interference against deubiquitinating enzymes can be used to elucidate their function. The application of this technology will greatly improve the functional annotation of this family of proteases.

Original publication




Journal article


Methods Enzymol

Publication Date





554 - 567


Cell Line, Tumor, Endopeptidases, Gene Library, Genetic Vectors, Humans, Multigene Family, RNA Interference, Substrate Specificity, Ubiquitin