[Construction of HCV-producing cell model based on self-cleaving ribozyme]

Area of Science:

• Virology
• Molecular Biology
• Cell Biology

Context:

• Hepatitis C virus (HCV) infection remains a significant global health challenge.
• Developing effective antiviral therapies requires robust experimental models.
• Existing models may not fully recapitulate stable HCV production for drug screening.

Purpose:

• To engineer a stable cell line that continuously produces infectious Hepatitis C virus (HCV).
• To establish a reliable platform for evaluating the efficacy of anti-HCV drugs.
• To facilitate research into HCV replication mechanisms and antiviral strategies.

Summary:

• A recombinant plasmid (pJFH1-Rbz) encoding the HCV JFH-1 genome flanked by ribozymes was constructed.
• Transfection into HepG2 cells followed by G418 screening yielded a stable HCV-producing clone.
• HCV core protein expression, high viral RNA levels (10^6 copies/ml), and viral particles (55 nm) were confirmed; IFN-alpha treatment reduced viral RNA.

Impact:

• Provides a valuable tool for preclinical anti-HCV drug development.
• Enables high-throughput screening of antiviral compounds.
• Facilitates a deeper understanding of HCV pathogenesis and therapeutic interventions.