Hepacivirus ; Hepatitis C ; genetics ; Humans ; MicroRNAs ; genetics

Genetic Heterogeneity ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Replicon ; genetics

Genes, Viral ; genetics ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Recombination, Genetic

Adult ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Turkey

Hepacivirus ; genetics ; Hepatitis C ; diagnosis ; genetics ; therapy ; Humans ; Interleukins ; genetics ; Polymorphism, Single Nucleotide ; Prognosis

DNA, Catalytic ; genetics ; Gene Expression ; Genetic Therapy ; Hep G2 Cells ; Hepacivirus ; genetics ; Humans ; Ribosomes ; genetics ; Transfection

Apoptosis ; Cell Line ; Hepacivirus ; genetics ; Humans ; Serum ; metabolism ; Viral Nonstructural Proteins ; genetics

Genome, Viral ; Hepacivirus ; genetics ; physiology ; Models, Biological ; RNA, Viral ; biosynthesis ; genetics ; Replicon ; Transformation, Genetic ; Virus Replication

Infection with hepatitis C virus (HCV) is one of the major global health problems, approximately 170 million people are infected worldwide. The chronic HCV infection is associated with a high risk of developing liver cirrhosis, hepatocellular carcinoma (HCC) and liver failure. Unfortunately, there is still no effective vaccine or antibodies available for the prevention of infection. RNA interference (RNAi) represents a promising new approach to combat viral infections, and recent developments in the field of gene therapy have increased the feasibility of clinical applications. RNAi techniques have made rapid progress in the basic understanding of HCV biology and revealed numerous new viral and host-cell factors as potential targets for therapy. Together with the improvement of gene delivery technique and the discovery of the critical role of microRNA (miRNA) in HCV infection, RNAi and miRNA-based antiviral strategies hold great promise for the future. In this article, we provide a comprehensive overview of current developments of therapeutic targets of RNAi, liver-targeted delivery systems and the potential applications of miRNAs in treatment of hepatitis C infection.
Genetic Therapy ; Hepacivirus ; genetics ; Hepatitis C ; therapy ; Humans ; MicroRNAs ; genetics ; RNA Interference

OBJECTIVETo study the cleavage activity of specific deoxyribozyme to hepatitis C virus in vitro.

METHODSThree deoxyribozymes were designed to cleave at sites 157, 168, 173 in HCV 5'-noncoding region with the active region of 5'-GGCTAGCTACAACGA-3' respectively. Plasmid pCMV/T7-NCRC -Delta Luc was completely linearized with restriction endonuclease Xba I. HCV RNA5'-NCRC was transcribed in vitro from the linearized products and radiolabelled with [alpha-32P] UTP. Under the conditions of 37 degrees C, pH7.5, Mg2+ 10 mmol/L, the three deoxyribozymes were mixed with substrate RNA individually for 120 minutes and then the reactions were terminated. The cleavaged products were separated with 8% denaturated polyacrylamide gel electrophoresis and displayed by autoradiography. DRz3 was mixed with the substrate RNA at different Mg2+ concentrations. The cleavage efficiency was analyzed with a gel document action analyzing systems.

RESULTSUnder the adopted conditions the three deoxyribozymes efficiently cleaved to the target RNA in vitro and the cleavage activity of DRz3 was increased with the increase of Mg2+ concentration.

CONCLUSIONThe designed deoxyribozymes can cleave 5'-NCR mRNA of HCV efficiently in vitro and it is dose-respondent to Mg2+ concentration.

DNA, Catalytic ; genetics ; DNA, Single-Stranded ; genetics ; Genetic Therapy ; Hepacivirus ; genetics ; Hepatitis C ; therapy ; Humans ; RNA, Messenger ; genetics