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Journal of Virology, February 2000, p. 2046-2051, Vol. 74, No. 4
Department of Molecular Microbiology,
Washington University School of Medicine, St. Louis, Missouri
63110-1093,1 and Division of Virology,
Center for Biologics Evaluation and Research, Food and Drug
Administration, Bethesda, Maryland 208922
Received 7 July 1999/Accepted 22 November 1999
Hepatitis C virus (HCV) infection is a widespread major human
health concern. Significant obstacles in the study of this virus include the absence of a reliable tissue culture system and a small-animal model. Recently, we constructed full-length HCV cDNA clones and successfully initiated HCV infection in two chimpanzees by
intrahepatic injection of in vitro-transcribed RNA (A. A. Kolykhalov et al., Science 277:570-574, 1997). In order to validate
potential targets for development of anti-HCV therapeutics, we
constructed six mutant derivatives of this prototype infectious clone.
Four clones contained point mutations ablating the activity of the NS2-3 protease, the NS3-4A serine protease, the NS3 NTPase/helicase, and the NS5B polymerase. Two additional clones contained deletions encompassing all or part of the highly conserved 98-base sequence at
the 3' terminus of the HCV genome RNA. The RNA transcript from each of
the six clones was injected intrahepatically into a chimpanzee. No
signs of HCV infection were detected in the 8 months following the
injection. Inoculation of the same animal with nonmutant RNA transcripts resulted in productive HCV infection, as evidenced by
viremia, elevated serum alanine aminotransferase, and HCV-specific seroconversion. These data suggest that these four HCV-encoded enzymatic activities and the conserved 3' terminal RNA element are
essential for productive replication in vivo.
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Hepatitis C Virus-Encoded Enzymatic Activities and
Conserved RNA Elements in the 3' Nontranslated Region Are Essential for
Virus Replication In Vivo
*
Corresponding author. Mailing address: Department of
Molecular Microbiology, Campus Box 8230, Washington University School of Medicine, 660 S. Euclid Ave., St. Louis, MO 63110-1093. Phone: (314)
362-2842. Fax: (314) 362-1232. E-mail:
rice{at}borcim.wustl.edu.
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