Fluorescent Nanosensor for Drug Discovery

Abstract

There is a pressing need for developing new anti-HIV agents due to the emergence of drug-resistant HIV mutants, side effects of existing drugs, and the mutation of the virus. High-throughput screening (HTS) has been proven as a powerful technique for the discovery of new anti-HIV drugs. The utilization of high-content screening (HCS) requires development of nanosensor that is suitable for HCS. We developed fluorescence imaging-based nanosensor for screening of inhibitors against activity of HIV-1 protease. We explored using AcGFP1 (a fluorescent mutant of the wild-type green fluorescent protein) and mCherry (a mutant of red fluorescent protein), as two fluorophores for Förster resonance energy transfer (FRET) microscopy imaging measurement of HIV-1 protease activity within living cells. Both in vitro and in vivo studies revealed that the novel molecular probes exhibit significant enhancement of FRET signals. The probe developed in this study enables HCS of new anti-HIV agents.