Elsevier

SLAS Discovery

Volume 16, Issue 2, February 2011, Pages 211-220
SLAS Discovery

Original Articles
Optimized High-Throughput Screen for Hepatitis C Virus Translation Inhibitors

https://doi.org/10.1177/1087057110391665Get rights and content
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Hepatitis C virus (HCV) is a considerable global health problem for which new classes of therapeutics are needed. The authors developed a high-throughput assay to identify compounds that selectively block translation initiation from the HCV internal ribosome entry site (HCV IRES). Rabbit reticulocyte lysate conditions were optimized to faithfully report on authentic HCV IRES-dependent translation relative to a 5′ capped mRNA control. The authors screened a library of ~430,000 small molecules for IRES inhibition, leading to ~1700 initial hits. After secondary counterscreening, the vast majority of hits proved to be luciferase and general translation inhibitors. Despite well-optimized in vitro translation conditions, in the end, the authors found no selective HCV IRES inhibitors but did discover a new scaffold of general translation inhibitor. The analysis of these molecules, as well we the finding that a large fraction of false positives resulted from off-target effects, highlights the challenges inherent in screens for RNA-specific inhibitors.

Key words

hepatitis C virus (HCV)
IRES
luciferase
high-throughput screen
rabbit reticulocyte lysate

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