[PDF] Top 20 Identification of the Hepatitis C Virus RNA Replication Complex in Huh-7 Cells Harboring Subgenomic Replicons

... First, Huh-7-derived clone 9-13, harboring a bicistronic subgenomic replicon (19), was investigated by immunofluores- cence microscopy (IF) to determine the subcellular distribu- tion of the ... See full document

... mutant replicons suggest that the coelectroporation method used to enumerate the levels of resistance within an RNA population may underestimate the actual numbers of replicons carrying point ...of ... See full document

... labeled replication products were eluted from an agarose gel and used as a probe for hybridization in a Northern blot which included the following RNAs ...replicon RNA (lane 1), an N-terminal por- tion of ... See full document

... HCV replicons in cell culture [9-11,20- 22], we sequenced replicon clones obtained from our mouse cell lines to determine whether the population of sgJFH-1 replicons present in the MMH cells had ... See full document

... with cells transduced with Luc-core ⌬ 212-NS2, Luc-no- core-NS2, and Luc-nocore-NS3 increased to levels higher than those observed at 5 h ...fected RNA. Replication of the RNA constructs at 72 ... See full document

... HCV RNA replication, the formation of foci containing dsRNA during the transient replication of the JFH-1 replicon in the presence of ribavirin, which is an inhibitor of HCV genome synthesis (24), ... See full document

... our subgenomic replicons do not express HCV core, making interpretation of the importance of the NS5A-core interaction impossible in our ...HCV replication in nonhepatic cell lines of human and mouse ... See full document

... technology. Huh-7 cells were transfected with siRNA pools targeting each of the BFA-sensitive ARF GEFs: brefeldin A- inhibited guanine nucleotide exchange factor 1 (BIG1), BIG2, or Golgi BFA ... See full document

... strand RNA) or was destroyed during cell lysis (20% of nega- tive-strand, 30% of positive-strand RNA) or during centrifu- gation of CRCs (15% of negative-strand and positive-strand RNA, ... See full document

... HCV replication, irrespective of whether IRF-3 was ex- pressed or ...HCV replication was still enhanced significantly in IRF-3-deficient mouse fibroblasts compared to that of the same replicon in wild-type ... See full document

... active Huh-7 proteome during HCV .../5.1) subgenomic replicon used in this study (NS, non-structural ...control) RNA (upper two blots) and detection of HCV NS3, HCV NS5A, and b -tubulin (as a ... See full document

... The hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp), represented by nonstructural protein 5B (NS5B), belongs to a class of integral membrane proteins termed tail-anchored ... See full document

... pSGR-JFH1 RNA and a nonreplicative pSGR-JFH1 RNA containing a lethal lesion in the RNA-dependent RNA polymerase (Pol ⫺ ) ...other replicons of different HCV genotypes, replicated ... See full document

... hepatoma cells, we examined whether IL-1 antiviral activity is associated with the activation of ERK MAP ...Huh7 cells and the replicon FCA1 cells were treated with 10 ng of IL-1␤ per ml for 30 min, ... See full document

... of Huh-7 cells permissive for HCV replication, we needed another line of Huh-7 cells also permissive for HCV replication but different from HRP1 ...transfected ... See full document

... single-plus-strand virus RNA genome ...single-stranded RNA viruses ...eukaryotic cells, developed additional mechanisms for efficient RNA amplification by co- localization of their ... See full document

... non-transfected cells, the total proteins were prepared by RIPA cell ...antigen-antibody complex was detected by an enhanced chemiluminescence (ECL) kit following the manufacturer ’ s ...in ... See full document

... unwind RNA, even though they retain some ability to unwind DNA and retain the same level of nucleic acid stimulated- ATPase activity as the wild type ...of cells containing replicons with R393A, ... See full document

... for replication of the full- length constructs containing the NS5A S2204I adaptive change ...In Huh-7.5 cells, the bicistronic construct containing the C-Neo cistron [FL-Neo (S2204I)] ...the ... See full document

... The current most effective therapy for HCV consists of pegylated interferon-α (IFN-α), administered once weekly plus daily oral ribavirin (RBV) for 24 to 48 weeks [11,12]. This combination therapy is quite successful in ... See full document