Therefore, CLDN and hCD81 were exogenously expressed in the cell lines, and the resulting lines were designated Huh6/CLDN and HepG2/CD81, respectively
Therefore, CLDN and hCD81 were exogenously expressed in the cell lines, and the resulting lines were designated Huh6/CLDN and HepG2/CD81, respectively. by removal of HCV RNA from your Hep3B/miR122 replicon cells, exhibited an enhanced expression of miR122 and a continuous increase of infectious titers of HCVcc in every passage. The Mouse monoclonal to CHK1 establishment of the new permissive cell collection for HCVcc will have significant implications not only for basic HCV research but also for the development of new therapeutics. INTRODUCTION Hepatitis C computer BAMB-4 virus (HCV) infects over 170 million people worldwide and frequently prospects to persistent contamination, which in turn can lead to chronic hepatitis, cirrhosis, and hepatocellular carcinoma (34). HCV belongs to the family and has a single-stranded positive RNA genome of approximately 9.6 kb. The genome of HCV is usually translated into a single polyprotein at the endoplasmic reticulum (ER) membrane and is then cleaved by host- and virus-encoded proteases, resulting in 10 structural and nonstructural proteins (41, 44). Due to the lack of a small-animal model and an efficient cell culture system, efforts to understand the HCV life cycle as well as development of anti-HCV drugs have been hampered (42). In a major breakthrough, HCV replicon cells, in which HCV RNA autonomously replicates, were established by Lohmann et al. (37). Afterwards, the infectious HCV in cell culture (HCVcc), based on the genotype 2a JFH1 strain in combination with the human hepatocellular carcinoma cell collection Huh7, was developed (36, 64, 70). On the basis of the results obtained with these BAMB-4 systems, the life cycle of HCV was clarified, and substantial progress has been made in screening host factors involved in HCV propagation as well as anti-HCV drug candidates (20, 51). Among them, a liver-specific microRNA (miRNA), miR122, has been shown to be one of the most important host factors for HCV replication. miRNAs are small noncoding RNAs that consist of 20 to 25 nucleotides and modulate gene expression in plants and animals (3, 26). Most miRNAs negatively regulate translation through conversation with the 3 untranslated region (UTR) of mRNA in a BAMB-4 sequence-specific manner. Some of them have been BAMB-4 shown to play important functions in the viral life cycle (56). Interestingly, miR122 has been shown to bind to HCV 5 UTRs and to enhance translation and replication of HCV RNA (23, 28, 29, 38, 52). In addition, enhancement of HCVcc propagation through the direct conversation of miR122 with HCV 5 UTR has been demonstrated (27). Recently, intravenous administration of the locked nucleic acid (LNA) complementary to miR122 was shown to suppress the propagation of HCV in chimpanzees chronically infected with HCV, suggesting that miR122 is usually a promising therapeutic target for chronic hepatitis C (31). It has been shown that HCV exploits numerous host factors to form a replication complex for efficient replication (43). propagation of HCV is limited to Huh7 cells and their derivatives, and thus, it is important to confirm the data obtained in Huh7 cells by using other human liver cell lines, because the patterns of gene expression vary among cell lines. Although establishment of an HCV replicon system based on liver cell lines has been reported (11, 66), strong propagation of HCVcc in well-characterized human liver cell lines other than Huh7 cells has not succeeded yet. The gene expression profile of mice xenotransplanted with human hepatocytes from different donors inoculated with a single source.