Engineering and Characterizing Synthetic Protease Sensors and Switches

Proteases are finding an increasing number of applications as molecular tools and reporters in biotechnology and basic research. Proteases are also increasingly incorporated into synthetic genetic signaling circuits equipping cells with tailored new functions. In the majority of cases however, proteases are employed in constitutively active forms which limits their utility and application as molecular sensors. The following chapter provides a detailed experimental protocol for converting constitutively active proteases into regulated protease receptors. Such receptors can potentially sense, transduce, and amplify any molecular input, thereby opening up a range of new applications in basic research, biotechnology, and synthetic biology.