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Inducible expression of human hepatitis B virus (HBV) in stably transfected hepatoblastoma cells: a novel system for screening potential inhibitors of HBV replication

作者:Stephanie K. Ladner, Michaël Otto, Christopher S. Barker, Katie Zaifert, G H Wang, Ju‐Tao Guo, Christoph Seeger, Robert W. King · 发表于:Antimicrobial Agents and Chemotherapy · 年份:1997 · DOI:10.1128/aac.41.8.1715 · 被引用次数:647 · 研究领域:Hepatitis B Virus Studies、RNA Interference and Gene Delivery、Virus-based gene therapy research

We report the development and isolation of a cell line, termed HepAD38, that replicates human hepatitis B virus (HBV) under conditions that can be regulated with tetracycline. In the presence of the antibiotic, this cell line is free of virus due to the repression of pregenomic (pg) RNA synthesis. Upon removal of tetracycline from the culture medium, the cells express viral pg RNA, accumulate subviral particles in the cytoplasm that contain DNA intermediates characteristic of viral replication, and secrete virus-like particles into the supernatant. Since the HepAD38 cell line can produce high levels of HBV DNA, it should be useful for analyses of the viral replication cycle that depend upon viral DNA synthesis in a synchronized fashion. In addition, this cell line has been formatted into a high-throughput, cell-based assay that permits the large-scale screening of diverse compound libraries for new classes of inhibitors of HBV replication.