Creation and characterization of a cell-death reporter cell line for hepatitis C virus infection. Academic Article uri icon

abstract

  • The present study describes the creation and characterization of a hepatoma cell line, n4mBid, that supports all stages of the hepatitis C virus (HCV) life cycle and strongly reports HCV infection by a cell-death phenotype. The n4mBid cell line is derived from the highly HCV-permissive Huh-7.5 hepatoma cell line and contains a modified Bid protein (mBid) that is cleaved and activated by the HCV serine protease NS3-4A. N4mBid exhibited a 10-20-fold difference in cell viability between the HCV-infected and mock-infected states, while the parental Huh-7.5 cells showed <2-fold difference under the same conditions. The pronounced difference in n4mBid cell viability between the HCV- and mock-infected states in a 96-well plate format points to its usefulness in cell survival-based high-throughput screens for anti-HCV molecules. The degree of cell death was found to be proportional to the intracellular load of HCV. HCV-low n4mBid cells, expressing an anti-HCV short hairpin RNA, showed a significant growth advantage over nave cells and could be rapidly enriched after HCV infection, suggesting the possibility of using n4mBid cells for the cell survival-based selection of genetic anti-HCV factors.

published proceedings

  • Antiviral Res

author list (cited authors)

  • Chen, Z., Simeon, R. L., Chockalingam, K., & Rice, C. M.

citation count

  • 9

complete list of authors

  • Chen, Zhilei||Simeon, Rudo L||Chockalingam, Karuppiah||Rice, Charles M

publication date

  • January 2010