HCV IRES-Mediated Core Expression in Zebrafish
- Authors
- Zhao, Y., Qin, W., Zhang, J.P., Hu, Z.Y., Tong, J.W., Ding, C.B., Peng, Z.G., Zhao, L.X., Song, D.Q., and Jiang, J.D.
- ID
- ZDB-PUB-130402-8
- Date
- 2013
- Source
- PLoS One 8(3): e56985 (Journal)
- Registered Authors
- Zhang, Jing-pu
- Keywords
- Larvae, Zebrafish, Hepatitis C virus, Internal ribosome entry site, Cobalamins, Reverse transcriptase-polymerase chain reaction, Gene expression, Liver fibrosis
- MeSH Terms
-
- Animals
- Antiviral Agents/pharmacology
- Biomarkers/metabolism
- Disease Models, Animal
- Dose-Response Relationship, Drug
- Gene Expression Regulation, Viral/drug effects*
- Genes, Reporter
- Green Fluorescent Proteins
- Hepacivirus/drug effects*
- Hepacivirus/genetics*
- Hepacivirus/growth & development
- Hepatitis C/drug therapy*
- Hepatitis C/virology
- Host-Pathogen Interactions
- Humans
- Interferon-alpha/pharmacology
- Larva/drug effects
- Larva/virology
- Liver/drug effects
- Liver/virology
- Mutant Chimeric Proteins/antagonists & inhibitors
- Mutant Chimeric Proteins/genetics
- Mutant Chimeric Proteins/metabolism
- RNA, Messenger/antagonists & inhibitors*
- RNA, Messenger/genetics
- Recombinant Proteins/pharmacology
- Ribavirin/pharmacology
- Viral Core Proteins/antagonists & inhibitors
- Viral Core Proteins/genetics*
- Viral Core Proteins/metabolism
- Vitamin B 12/pharmacology
- Zebrafish/virology*
- PubMed
- 23469178 Full text @ PLoS One
The lack of small animal models for hepatitis C virus has impeded the discovery and development of anti-HCV drugs. HCV-IRES plays an important role in HCV gene expression, and is an attractive target for antiviral therapy. In this study, we report a zebrafish model with a biscistron expression construct that can co-transcribe GFP and HCV-core genes by human hepatic lipase promoter and zebrafish liver fatty acid binding protein enhancer. HCV core translation was designed mediated by HCV-IRES sequence and gfp was by a canonical cap-dependent mechanism. Results of fluorescence image and in situ hybridization indicate that expression of HCV core and GFP is liver-specific; RT-PCR and Western blotting show that both core and gfp expression are elevated in a time-dependent manner for both transcription and translation. It means that the HCV-IRES exerted its role in this zebrafish model. Furthermore, the liver-pathological impact associated with HCV-infection was detected by examination of gene markers and some of them were elevated, such as adiponectin receptor, heparanase, TGF-β, PDGF-α, etc. The model was used to evaluate three clinical drugs, ribavirin, IFNα-2b and vitamin B12. The results show that vitamin B12 inhibited core expression in mRNA and protein levels in dose-dependent manner, but failed to impact gfp expression. Also VB12 down-regulated some gene transcriptions involved in fat liver, liver fibrosis and HCV-associated pathological process in the larvae. It reveals that HCV-IRES responds to vitamin B12 sensitively in the zebrafish model. Ribavirin did not disturb core expression, hinting that HCV-IRES is not a target site of ribavirin. IFNα-2b was not active, which maybe resulted from its degradation in vivo for the long time. These findings demonstrate the feasibility of the zebrafish model for screening of anti-HCV drugs targeting to HCV-IRES. The zebrafish system provides a novel evidence of using zebrafish as a HCV model organism.