This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zhang, Y.
Right arrow Articles by Hao, W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, Y.
Right arrow Articles by Hao, W.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, February 2008, p. 666-674, Vol. 52, No. 2
0066-4804/08/$08.00+0     doi:10.1128/AAC.01133-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Novel Chimeric Genotype 1b/2a Hepatitis C Virus Suitable for High-Throughput Screening{triangledown} ,{dagger}

Yingjia Zhang, Peter Weady, Rohit Duggal, and Weidong Hao*

Department of Exploratory Biology, Pfizer Global Research and Development, La Jolla laboratories, 10628 Science Center Drive, San Diego, California 92121

Received 27 August 2007/ Returned for modification 9 October 2007/ Accepted 12 November 2007

A major obstacle in hepatitis C virus (HCV) research has been the lack of a permissive cell culture system that produces infectious viral particles. Significant breakthroughs have been achieved lately in establishing such culture systems. Yet to date, there are no reports of the applications of any of these systems in HCV drug screening. Here, we report the generation of two monocistronic, chimeric genotype 1 full-length HCV genome molecules. These molecules, C33J-Y835C-UBI and C33J-Y835C-FMDV2A, both contain the structural protein region from genotype 1 (subtype 1b, Con1) and the remaining region from the genotype 2a (JFH1) clone. Both contain the humanized Renilla luciferase reporter gene which is separated from the rest of the HCV open reading frame by two different cleavage sites. The viral RNAs replicated efficiently in transfected cells. Viral particles produced were infectious in naïve Huh7.5 cells, and the infectivity could be blocked by monoclonal antibody against a putative HCV entry cofactor, CD81. A pilot high-throughput screen of 900 unknown compounds was executed by both the genotype 2a subgenomic replicon system and the infectious system. Thirty-one compounds were identified as hits by both systems, whereas 78 compounds were identified as hits only for the infectious system, suggesting that the infectious system is capable of identifying inhibitors targeting the viral structural proteins and steps involving them in the viral life cycle. The infectious HCV system developed here provides a useful and versatile tool which should greatly facilitate the identification of HCV inhibitors currently not identified by the subgenomic replicon system.

* Corresponding author. Mailing address: Department of Exploratory Biology, Pfizer Global Research and Development, La Jolla Laboratories, 10628 Science Center Drive, San Diego, CA 92121. Phone: (858) 638-3795. Fax: (858) 526-4502. E-mail: weidong.hao{at}pfizer.com

{triangledown} Published ahead of print on 26 November 2007.

{dagger} Supplemental material for this article may be found at http://aac.asm.org/.

Antimicrobial Agents and Chemotherapy, February 2008, p. 666-674, Vol. 52, No. 2
0066-4804/08/$08.00+0     doi:10.1128/AAC.01133-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Pan, K.-L., Lee, J.-C., Sung, H.-W., Chang, T.-Y., Hsu, J. T.-A. (2009). Development of NS3/4A Protease-Based Reporter Assay Suitable for Efficiently Assessing Hepatitis C Virus Infection. Antimicrob. Agents Chemother. 53: 4825-4834 [Abstract] [Full Text]  
  • Yu, X., Sainz, B. Jr., Uprichard, S. L. (2009). Development of a Cell-Based Hepatitis C Virus Infection Fluorescent Resonance Energy Transfer Assay for High-Throughput Antiviral Compound Screening. Antimicrob. Agents Chemother. 53: 4311-4319 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»