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Infectious cDNA clone of the hepatitis C virus genotype 1 prototype sequence
Robert E Lanford1, Helen Lee1, Deborah Chavez1
1Department of Virology and Immunology1 and Department of Laboratory Animal Medicine2, Southwest Regional Primate Research Center, Southwest Foundation for Biomedical Research, 7620 NW Loop 410, San Antonio, TX 78227, USA.
The Journal of General Virology
|May 23, 2001
Summary
Researchers created an infectious clone of the hepatitis C virus (HCV) prototype sequence. This breakthrough enables further study of HCV genotype 1a, crucial for developing diagnostics and therapeutics.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- The hepatitis C virus (HCV) is a significant global health concern.
- Developing infectious molecular clones is essential for understanding HCV replication and pathogenesis.
- The HCV genotype 1 prototype sequence (HCV-1) is widely used but lacked a functional infectious clone.
Purpose of the Study:
- To construct a full-length infectious cDNA clone of the HCV genotype 1 prototype (subtype 1a).
- To generate an infectious chimeric HCV clone for further research.
Main Methods:
- Construction of a full-length cDNA clone of HCV genotype 1a.
- Preparation of a consensus clone by comparing multiple HCV sequences.
- Site-directed mutagenesis to introduce 11 non-consensus amino acid changes.
- In vitro synthesis of RNA and intrahepatic inoculation of chimpanzees.
- Construction and testing of chimeric cDNA clones between HCV-1 and H77 strains.
Main Results:
- The initial HCV-1 cDNA clone did not produce infectious virus.
- Synthetic RNA from the repaired consensus clone initiated a typical acute-resolving HCV infection in a chimpanzee.
- At least one of three chimeric cDNA clones between HCV-1 and H77 was infectious in a chimpanzee.
- This represents the first infectious chimeric HCV clone.
Conclusions:
- An infectious cDNA clone of the HCV genotype 1 prototype (HCV-1) has been successfully constructed.
- This infectious clone is invaluable for research, given its status as the prototype sequence.
- The development of infectious chimeric clones opens new avenues for HCV research.