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Journal of Biomolecular Screening
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Assay Optimization and Screening of RNA-Protein Interactions by AlphaScreen

Nicholas L. Mills

Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee

Anang A. Shelat

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee

R. Kiplin Guy

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee, Kip.Guy{at}StJude.org

The lack of lead compounds that specifically recognize and manipulate the function of RNA molecules limits our ability to consider RNA targets valid for drug discovery. Herein is reported a high-throughput biochemical screen for inhibitors of RNA-protein interactions based on AlphaScreen technology that incorporates several layers of specificity measurements into the primary screen. This screen was used to analyze approximately 5500 compounds from a collection of bioactive small molecules to detect inhibitors of the HIV-1 Rev-RRE and BIV Tat-TAR interactions. This proof-of-concept screen validates the assay as one that accurately identifies hit molecules and determines the selectivity of those hits. (Journal of Biomolecular Screening 2007: 946-955)

Key Words: HIV-1 Rev • Rev-responsive element • bovine immunodeficiency virus • fluorescence polarization

Journal of Biomolecular Screening, Vol. 12, No. 7, 946-955 (2007)
DOI: 10.1177/1087057107306128
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[Abstract] [PDF]