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Inhibition of hepatitis C virus replication by single-stranded RNA structural mimics

World Journal of Gastroenterology 2010年第17期16卷 2100-2108页

作者： Robert Smolic Martina Smolic John H Andorfer Catherine H Wu Robert M Smith George Y Wu Department of Medicine Division of Gastroenterology-HepatologyUniversity of Connecticut Health Center263 Farmington Ave.FarmingtonCT 06030-1845United States

AIM: To examine the effect of hepatitis C virus (HCV) structural mimics of regulatory regions of the genome on HCV ***: HCV RNA structural mimics were constructed and tested in a HCV genotype 1b aBB7 replicon,and a japanese fulminant hepatitis-1 (JFH-1) HCV genotype 2a infection *** sequences were computer-predicted to adopt stem-loop structures identical to the corresponding elements in full-length viral ***7.5 cells bearing the BB7 replicon or infected with JFH-1 virus were transfected with expression vectors generating HCV mimics and *** HCV RNA and protein levels were quantified by real-time polymerase chain reaction and Western blotting,*** evaluate possible antisense effects,complementary RNAs spanning a mimic were ***: In the BB7 genotype 1b replicon system,mimics of the polymerase (NS-5B),X and BA regions inhibited replication by more than 90%,50%,and 60%,*** the JFH-1 genotype 2 infection system,mimics that were only 74% and 46% identical in sequence relative to the corresponding region in JFH-1 inhibited HCV replication by 91.5% and 91.2%,respectively,as effectively as a mimic with complete identity to HCV genotype *** inhibitory effects were confirmed by NS3 protein *** RNA molecules spanning the 74% identical mimic had no significant ***: HCV RNA structural mimics can inhibit HCV RNA replication in replicon and infectious HCV systems and do so independent of close sequence identity with the target.

关键词： hepatitis C virus japanese fulminant hepatitis virus Complementarity RNA sequence Hybridization

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Secondary Structural Elements of the HCV X-region Involved in Viral Replication

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Journal of Clinical and Translational Hepatology 2015年第1期3卷 1-8页

作者： Nidhi Gupta Catherine H.Wu George Y.Wu Department of Medicine Division of Gastroenterology-Hepatology University of Connecticut Health Center Farmington CT USA

Background and Aims:The noncoding regions in the 3'-untranslated region (UTR) of the hepatitis C virus (HCV)genome contain secondary structures that are important for *** aim of this study was to identify detailed conformational elements of the X-region involved in HCV ***:Ribonucleic acid (RNA) structural analogs X94,X12,and X12c were constructed to have identical conformation but 94%,12%,and 0% sequence identity,respectively,to the X region of HCV genotype *** of structural analogs on replication of HCV genotypes 1b and 2a HCV RNA were studied by quantitative reverse transcriptase polymerase chain ***:In replicon BB7 cells,a constitutive replication model,HCV RNA levels decreased to 55%,52%,53%,and 54% after transfection with expression plasmids generating RNA structural analogs 5B-46,X-94,X-12,and X-12c,respectively (p<0.001 for all).In an HCV genotype 2a infection model,RNA analogs 5B-46,X-94,and X-12 in hepatic cells inhibited replication to 11%,9%,and 12%,*** the X-12 analog was only 12% identical to the corresponding sequence of HCV genotype 2a,the sequence per se,or antisense effects were unlikely to be ***:The data suggest that conformation of secondary structures in 3'-UTR of HCV RNA genome is required for HCV *** expression of RNA analogs predicted to have identical stem-loop structures might inhibit HCV infection of hepatocytes in liver and may represent a novel approach to design anti-HCV agents.

关键词： HCV RNA genome HCV X-region hepatitis C virus Infection japanese fulminant hepatitis virus RNA secondary structure

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