title: Zebrafish as a potential model organism for drug test against hepatitis C virus.
abstract: Screening and evaluating anti- hepatitis C virus (HCV) drugs in vivo is difficult worldwide, mainly because of the lack of suitable small animal models. We investigate whether zebrafish could be a model organism for HCV replication. To achieve NS5B-dependent replication an HCV sub-replicon was designed and created with two vectors, one with HCV ns5b and fluorescent rfp genes, and the other containing HCV's 5'UTR, core, 3'UTR and fluorescent gfp genes. The vectors containing sub-replicons were co-injected into zebrafish zygotes. The sub-replicon amplified in liver showing a significant expression of HCV core RNA and protein. The sub-replicon amplification caused no abnormality in development and growth of zebrafish larvae, but induced gene expression change similar to that in human hepatocytes. As the amplified core fluorescence in live zebrafish was detectable microscopically, it rendered us an advantage to select those with replicating sub-replicon for drug experiments. Ribavirin and oxymatrine, two known anti-HCV drugs, inhibited sub-replicon amplification in this model showing reduced levels of HCV core RNA and protein. Technically, this method had a good reproducibility and is easy to operate. Thus, zebrafish might be a model organism to host HCV, and this zebrafish/HCV (sub-replicon) system could be an animal model for anti-HCV drug screening and evaluation.
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
authors.forename: Cun-Bao
authors.initials: CB
authors.lastname: Ding
authors.forename: Jing-Pu
authors.initials: JP
authors.lastname: Zhang
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhao
authors.forename: Zong-Gen
authors.initials: ZG
authors.lastname: Peng
authors.forename: Dan-Qing
authors.initials: DQ
authors.lastname: Song
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Jiang
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D015153:Blotting, Western; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005393:Fish Diseases / Q000517:prevention & control / Q000821:virology; D005784:Gene Amplification / Q000187:drug effects; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D006526:Hepatitis C / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D006801:Humans; D017403:In Situ Hybridization; D007814:Larva / Q000235:genetics / Q000378:metabolism / Q000821:virology; D008845:Microinjections; D008856:Microscopy, Fluorescence; D011807:Quinolizines / Q000494:pharmacology; D012093:Replicon / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012254:Ribavirin / Q000494:pharmacology; D020506:Untranslated Regions / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D000093842:Matrines; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000470:Alkaloids; D000998:Antiviral Agents; D049452:Green Fluorescent Proteins; D011807:Quinolizines; D012254:Ribavirin; D020506:Untranslated Regions; D017361:Viral Nonstructural Proteins; C037573:oxymatrine; D000093842:Matrines; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase