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HCV IRES-mediated core expression in zebrafish.
PLoS One. 2013; 8(3):e56985.Plos

Abstract

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.

Authors+Show Affiliations

Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23469178

Citation

Zhao, Ye, et al. "HCV IRES-mediated Core Expression in Zebrafish." PloS One, vol. 8, no. 3, 2013, pp. e56985.
Zhao Y, Qin W, Zhang JP, et al. HCV IRES-mediated core expression in zebrafish. PLoS ONE. 2013;8(3):e56985.
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., & Jiang, J. D. (2013). HCV IRES-mediated core expression in zebrafish. PloS One, 8(3), e56985. https://doi.org/10.1371/journal.pone.0056985
Zhao Y, et al. HCV IRES-mediated Core Expression in Zebrafish. PLoS ONE. 2013;8(3):e56985. PubMed PMID: 23469178.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - HCV IRES-mediated core expression in zebrafish. AU - Zhao,Ye, AU - Qin,Wei, AU - Zhang,Jing-Pu, AU - Hu,Zhan-Ying, AU - Tong,Jun-Wei, AU - Ding,Cun-Bao, AU - Peng,Zong-Gen, AU - Zhao,Li-Xun, AU - Song,Dan-Qing, AU - Jiang,Jian-Dong, Y1 - 2013/03/01/ PY - 2012/02/02/received PY - 2013/01/17/accepted PY - 2013/3/8/entrez PY - 2013/3/8/pubmed PY - 2013/8/28/medline SP - e56985 EP - e56985 JF - PloS one JO - PLoS ONE VL - 8 IS - 3 N2 - 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. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/23469178/HCV_IRES_mediated_core_expression_in_zebrafish_ L2 - http://dx.plos.org/10.1371/journal.pone.0056985 DB - PRIME DP - Unbound Medicine ER -