2. Academic Validation
  3. A high-throughput approach to evaluating NCp7 RNA binding activity for HIV-1 drug discovery

A high-throughput approach to evaluating NCp7 RNA binding activity for HIV-1 drug discovery

  • SLAS Discov. 2025 Aug 11:35:100260. doi: 10.1016/j.slasd.2025.100260.
Joanna K Winstone 1,  Rikki Uhrich 2,  Thibault Alle 3,  Brian C Kraemer 4
Affiliations

Affiliations

  • 1 Geriatrics Research Education and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA; Division of Gerontology and Geriatric Medicine, Department of Medicine, University of Washington, Seattle, WA 98104, USA.
  • 2 Geriatrics Research Education and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA.
  • 3 Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, La Jolla, California 92093, USA.
  • 4 Geriatrics Research Education and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA; Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, La Jolla, California 92093, USA; Department of Psychiatry and Behavioral Sciences, University of Washington, Seattle, Washington 98195, USA; Department of Pathology, University of Washington, Seattle, Washington 98195, USA. Electronic address: kraemerb@u.washington.edu.
PMID: 40803428 DOI: 10.1016/j.slasd.2025.100260
Abstract

The HIV-1 epidemic broadly impacts healthcare. There remains a continued need for improved anti-viral therapies resilient to the development of drug resistance. HIV-1 nucleocapsid protein 7 (NCp7) seems a prime drug target due to its unique nucleic acid chaperone activity required for multiple viral processes. NCp7 RNA binding activity has been shown to increase viral production and infectivity within the host. Here we introduce a high-throughput AlphaScreen assay to evaluate NCp7 RNA binding activity and validate its specificity and sensitivity using a known inhibitor. We also demonstrate the utility of this assay by performing a drug-repurposing screen, which identified seven confirmed inhibitors of NCp7 RNA binding and two confirmed enhancers of NCp7 RNA binding. This tool will aid in future NCp7-targeted drug discovery initiatives for the treatment of HIV-1 Infection.

Keywords

AlphaLISA; Drug discovery; HIV; NCp7; Nucleic acid binding; U3.

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