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Abstract

Methods and compositions for detecting the presence of an active protease in a cell are provided. A feature of the subject methods is that a protease detection fusion protein is employed to detect the protease activity of interest. The protease detection fusion protein includes first and second subcellular localization domains separated by a protease cleavage domain, where the first subcellular localization domain is dominant over the second. The protease detection fusion proteins employed in the subject methods are further characterized by having a label domain located between the protease cleavage and second subcellular localization domains. In practicing the subject methods, the protease detection fusion protein is first provided inside the cell to be assayed. Following a suitable incubation period, the subcellular location of the label domain is determined, where the location is indicative of whether or not the protease activity of interest is present in the cell. Also provided are systems and kits for use in practicing the subject methods. The subject invention finds use in a variety of different applications, including protease activity detection applications, drug screening applications, etc.

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First Claim