A Genetically Programmable Module for Screening of Protease Inhibitors based on Quantum Dot-Donor Fluorescence Resonance Energy Transfer

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A Genetically Programmable Module for Screening of Protease Inhibitors Based on Quantum Dot-Donor Fluorescence Resonance Energy TransferWilfred Chen University of California-RiversideCBET-0755775Intellectual Merit The overall objective of this collaborative project is to develop a genetically programmable module that is easily adaptable for screening inhibitors for a wide range of proteases. The approach is to generate a quantum dot (QD)-modified, protease-specific protein module that can be used as a Fluorescence Resonance Energy Transfer (FRET) substrate for probing protease activity. This will result in a new generation of genetically programmable protein modules as QD-based FRET substrates for protease inhibitor discovery. High-throughput screening analysis (HTS) of protease activity will be achieved using these new QD-based FRET substrates that can be delivered intracellularly by a cell-penetrating TAT peptide. The combination will enable the Principal Investigators to provide a rapid and sensitive assessment of protease activity and the corresponding inhibitor efficacies in a HTS manner.Broader Impact: The integration of the development of tunable biomolecules with FRET-based HTS screening represents a unique effort that expands the fundamental development of protein engineering with the implementation of drug discovery. Graduate students and postdoctoral researchers participating in this research will gain an integrated perspective of the important interfaces and synergies connecting biochemistry, modern genetics, and HTS.