Hepatitis C Virus Subgenomic Replicons in the Human Embryonic Kidney 293 Cell Line

Abstract

Hepatitis C virus (HCV) infects liver cells and its replication in other cells

Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density lipoproteins

“…9). This result is consistent with previous findings that HCV RNA can replicate in HeLa and HEK-293 cells (31)(32)(33), which do not produce VLDL. The reason for colocalization of the HCV replication and VLDL assembly appears to lie in a requirement for coassembly or secretion of VLDL and HCV particles.…”

Iron Inactivates the RNA Polymerase NS5B and Suppresses Subgenomic Replication of Hepatitis C Virus

“…4). Whereas the initial results were obtained in a single replicon clone (24), the iron-mediated inhibition in replicon expression was recapitulated in newly established Huh7 replicon clones and in 293Rep cells (26) (Fig. 5).…”

“…Cell Culture-Replicon and parent human Huh7 hepatoma cells (24,25) and human embryonic kidney 293 cells (26) were cultured in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum, 1% non-essential amino acids, 100 units/ml penicillin, and 100 g/ml streptomycin. The replicon cells were maintained in media containing 500 g/ml G418 (Geneticin; Invitrogen) in addition to the above-mentioned supplements.…”

“…To further validate this interpretation, we utilized the recently described 293Rep replicon system, consisting of human embryonic kidney 293 cells fused with the S22.3 clone of replicon Huh7 cells into a heterokaryon (26). Parent 293 and replicon 293Rep cells were subjected to iron manipulations, and the expression of NS5A protein and replicon RNA was analyzed by Western and Northern blotting, respectively.…”

A novel function for a micro RNA: Negative regulators can do positive for the hepatitis C virus

“…The study by Jopling and colleagues clearly shows that HCV RNA replication can be modulated by the abundance of miR-122 in Huh-7 cells. However, miR-122 is not expressed in several other cell lines that also support HCV RNA replication such as HeLa, 21,22 HepG2, 19,20 or 293T cells 22,25 arguing that miR-122 is not a major determinant of host cell permissiveness and that miR-122-HCV RNA interaction per se is not required for viral RNA replication. However, it is possible that in HeLa or HepG2 cells carrying a stably replicating HCV RNA miR-122 levels are upregulated and in this way host cell permissiveness is enhanced.…”