Article abstract
Nature Biotechnology 26, 1019 - 1027 (2008)
Published online: 31 August 2008 | doi:10.1038/nbt.1490
Discovery of a hepatitis C target and its pharmacological inhibitors by microfluidic affinity analysis
Shirit Einav1,2,5, Doron Gerber3,5, Paul D Bryson2, Ella H Sklan2, Menashe Elazar2, Sebastian J Maerkl3,4, Jeffrey S Glenn2 & Stephen R Quake3
Abstract
More effective therapies are urgently needed against hepatitis C virus (HCV), a major cause of viral hepatitis. We used in vitro protein expression and microfluidic affinity analysis to study RNA binding by the HCV transmembrane protein NS4B, which plays an essential role in HCV RNA replication. We show that HCV NS4B binds RNA and that this binding is specific for the 3' terminus of the negative strand of the viral genome with a dissociation constant (Kd) of 
3.4 nM. A high-throughput microfluidic screen of a compound library identified 18 compounds that substantially inhibited binding of RNA by NS4B. One of these compounds, clemizole hydrochloride, was found to inhibit HCV RNA replication in cell culture that was mediated by its suppression of NS4B's RNA binding, with little toxicity for the host cell. These results yield new insight into the HCV life cycle and provide a candidate compound for pharmaceutical development.
- Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, California 94305, USA.
- Department of Medicine, Division of Gastroenterology and Hepatology, Stanford University School of Medicine, CCSR 3115A, 269 Campus Drive, Stanford, California 94305, USA.
- Department of Bioengineering, Stanford University and Howard Hughes Medical Institute, Stanford, California 94305, USA.
- Present address: School of Engineering, École Polytechnique Fédérale de Lausanne, Building BM 2111, Station 17, 1015 Lausanne, Switzerland.
- These authors contributed equally to this work.
Correspondence to: Jeffrey S Glenn2 e-mail: jeffrey.glenn@stanford.edu
Correspondence to: Stephen R Quake3 e-mail: quake@stanford.edu
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RESEARCH
An in vitro microfluidic approach to generating protein-interaction networksNature Methods Brief Communication (01 Jan 2009)
