Abstract: A proximity dependent split T7 RNAP (RNA polymerase) sensor using continuous molecular evolution is described. The versatility of the platform is described by creating robust light and small molecule-responsive genetic sensors. The activity-responsive RNAP platform dramatically simplifies and expands genetic circuit creation, and opens new opportunities in protein engineering, synthetic biology, and bioengineering.

Abstract: Aspects of the disclosure relate to a RNA regulatory system comprising at least one of each: i) a RNA hairpin binding domain; ii) a RNA targeting molecule comprising a RNA targeting region and at least one hairpin structure, wherein the hairpin structure of the RNA targeting molecule specifically binds to i; and iii) a RNA regulatory domain.

Abstract: A proximity dependent split T7 RNAP (RNA polymerase) sensor using continuous molecular evolution is described. The versatility of the platform is described by creating robust light and small molecule-responsive genetic sensors. The activity-responsive RNAP platform dramatically simplifies and expands genetic circuit creation, and opens new opportunities in protein engineering, synthetic biology, and bioengineering.

Abstract: Synthetic probes for detecting the activity of enzymes that catalyze reactions of post-translationally modified cysteine residues are described. The probes include “turn-on” probes that include a carbamate linkage that is cleaved via an intramolecular reaction with a free thiol produced by an enzyme catalyzed activity. The probes also include ratiometric, Michael addition-based probes that respond to enzymatic activity by a change in structure that results in a change in fluorescence properties. Methods of using the probes to detect enzymatic activity and disease are described. For example, the probes can be used to detect enzymatic activity in a variety of samples, including live cells and heterogeneous tissues. In addition, prodrugs that can be activated by enzymes that catalyze reactions of post-translationally modified cysteine residues and methods of using the prodrugs to treat disease are described.

Abstract: Synthetic probes for detecting the activity of enzymes that catalyze reactions of post-translationally modified cysteine residues are described. The probes include “turn-on” probes that include a carbamate linkage that is cleaved via an intramolecular reaction with a free thiol produced by an enzyme catalyzed activity. The probes also include ratiometric, Michael addition-based probes that respond to enzymatic activity by a change in structure that results in a change in fluorescence properties. Methods of using the probes to detect enzymatic activity and disease are described. For example, the probes can be used to detect enzymatic activity in a variety of samples, including live cells and heterogeneous tissues. In addition, prodrugs that can be activated by enzymes that catalyze reactions of post-translationally modified cysteine residues and methods of using the prodrugs to treat disease are described.