| GeneLibs

题目:: Visualizing the essential role of complete virion assembly machinery in efficient hepatitis C virus cell-to-cell transmission by viral infection activated split-intein mediated reporter system (VISI).
作者:: Zhao(Fanfan),Zhao(Ting),Deng(Libin),Lv(Dawei),Zhang(Xiaolong),Pan(Xiaoyu),Xu(Jun),Long(Gang)
状态:: 发布时间2016-11-17 , 更新时间 2016-11-18
期刊:: J Virol
摘要:: Hepatitis C virus (HCV) infects 2-3% of the world population and is a leading cause of liver diseases such as fibrosis, cirrhosis and hepatocellular carcinoma. Many aspects of HCV study, ranging from molecular virology, antiviral drug development and drug resistance profiling, were supported by straightforward assays of HCV replication and infection. Among these assays, HCV-dependent fluorescence relocalization (HDFR) system allows live-cell visualization of infection without modifying viral genome, but this strategy required careful recognition of fluorescence relocalization pattern for its high fluorescence background in cytoplasm. In this study, to achieve background free visualization of HCV infection, a viral infection activated split-intein mediated reporter system (VISI) was devised. Uninfected huh7.5.1-VISI cell shows no background signal, while HCV infection specifically illuminates the nuclei of infected Huh7.5.1-VISI cells with either GFP or mCherry. Combining VISI-GFP and VISI-mCherry systems, we revisited HCV cell-to-cell transmission with clear-cut distinction of donor and recipient cells in live-cell manner. Independent of virion assembly, exosomes have been reported to transfer HCV sub-genomic RNA to initiate replication in uninfected cells, which suggested an assembly free pathway. However, our data demonstrated that HCV structural genes and p7 gene were essential for not only cell-free infectivity but also cell-to-cell transmission. Additionally, depletion of apoE from donor cells but not from recipient cells significantly reduced HCV cell-to-cell transmission efficiency. In summary, we developed a background-free cell-based reporter system for convenient live-cell visualization of HCV infection and our data indicate that complete HCV virion assembly machinery is essential for both cell-free and cell-to-cell transmission.,Hepatitis C virus (HCV) infects hepatocytes via two pathways: cell-free infection and cell-to-cell transmission. Structural module of HCV genome are required for production of infectious cell-free virions, however role of specific genes within structural module in cell-to-cell transmission is not clearly defined. Our data demonstrate that deletion of core, E1E2 and p7 gene individually results in no HCV cell-to-cell transmission and apoE knockdown from donor cells causes less efficient cell-to-cell transmission. Thus this work indicates complete HCV assembly machinery is required for HCV cell-to-cell transmission. At last this work presents an optimized viral infection activated split-intein mediated reporter system for easy live-cell monitoring of HCV infection.
语言:: eng
DOI:: 10.1128/JVI.01720-16

文献库文献相关信息

DataTable pData

联系方式

微信公众号

文献库 文献相关信息

DataTable pData

联系方式

微信公众号

文献库文献相关信息