Merged screening for human immunodeficiency virus Tat and Rev inhibitors

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Conventional drug discovery primarily targets proteins, but gene regulation via protein-RNA interactions is emerging as a novel therapeutic concept.
• Human immunodeficiency virus (HIV) replication critically depends on two specific protein-RNA interactions: Tat-TAR and Rev-RRE.
• Targeting these interactions offers a new strategy for developing antiviral therapies.

Purpose of the Study:

• To establish a high-throughput screening assay for identifying inhibitors of essential protein-RNA interactions in HIV.
• To simultaneously evaluate the potency and selectivity of potential drug candidates against two distinct protein-RNA targets.
• To explore the potential of RNA-binding compounds in modulating cellular regulatory processes.

Main Methods:

• Development of a fluorescence-based, microwell-based dual screening assay.
• Simultaneous interrogation of two key protein-RNA interactions (Tat-TAR and Rev-RRE) within the same reaction vessel.
• Screening of over 110,000 diverse chemical compounds.

Main Results:

• Successful setup and validation of a high-throughput screening assay for RNA binders.
• Identification and primary characterization of hit compounds demonstrating interference with protein-RNA interactions.
• Demonstration of the assay's capability to assess both potency and selectivity of inhibitors.

Conclusions:

• A microwell-based dual screening system for RNA binders represents a powerful new tool for modern drug discovery.
• This assay facilitates the identification of novel inhibitors targeting critical protein-RNA interactions.
• The approach has the potential to advance the development of therapeutics by targeting gene regulation.