Small molecule inhibition of RISC loading.
Publication/Presentation Date
2-17-2012
Abstract
Argonaute proteins are the core components of the microRNP/RISC. The biogenesis and function of microRNAs and endo- and exo- siRNAs are regulated by Ago2, an Argonaute protein with RNA binding and nuclease activities. Currently, there are no in vitro assays suitable for large-scale screening of microRNP/RISC loading modulators. We describe a novel in vitro assay that is based on fluorescence polarization of TAMRA-labeled RNAs loaded to human Ago2. Using this assay, we identified potent small-molecule inhibitors of RISC loading, including aurintricarboxylic acid (IC(50) = 0.47 μM), suramin (IC(50) = 0.69 μM), and oxidopamine HCL (IC(50) = 1.61 μM). Small molecules identified by this biochemical screening assay also inhibited siRNA loading to endogenous Ago2 in cultured cells.
Volume
7
Issue
2
First Page
403
Last Page
410
ISSN
1554-8937
Published In/Presented At
Tan, G. S., Chiu, C. H., Garchow, B. G., Metzler, D., Diamond, S. L., & Kiriakidou, M. (2012). Small molecule inhibition of RISC loading. ACS chemical biology, 7(2), 403–410. https://doi.org/10.1021/cb200253h
Disciplines
Business Administration, Management, and Operations | Health and Medical Administration | Management Sciences and Quantitative Methods
PubMedID
22026461
Department(s)
Administration and Leadership
Document Type
Article