Abstract
The 5′ untranslated region (5′ UTR) of rodent hepacivirus (RHV) and pegivirus (RPgV) contains sequence homology to the HCV type III internal ribosome entry sites (IRES). Utilizing a monocistronic expression vector with an RNA polymerase I promoter to drive transcription, we show cell-specific IRES translation and regions within the IRES required for full functionality. Focusing on RHV, we further pseudotyped lentivirus with RHV and showed cell surface expression of the envelope proteins and transduction of murine hepatocytes and we then constructed full-length RHV and RPgV replicons with reporter genes. Using the replicon system, we show that the RHV NS3-4A protease cleaves a mitochondrial antiviral signaling protein reporter. However, liver-derived cells did not readily support the complete viral life cycle.
Highlights
Introduction e hepatitisC virus (HCV) infects more than 71 million people worldwide [1], which can lead to liver failure and hepatocellular carcinoma and presents a major global health burden
Polymerase I promoter in front of the full-length viral 5′ untranslated region (5′ untranslated regions (UTRs)) followed by a fluorescent maker, the viral 3′ UTR, and the RNA polymerase I terminator (Figure 2(a)). e hepatitisC virus (HCV) internal ribosome entry sites (IRES) was used as a positive control along with a control plasmid containing a scrambled sequence in place of the viral 5′ UTR
Ese plasmids were transfected into the murine hepatocyte cell line Hepa1-6, with the HCV and rodent pegivirus (RPgV) IRES driving the highest level of translation at 72 hours after transfection with a mean fluorescence intensity (MFI) of 30, followed by RHV1 and RHV2 with an MFI of 22 and 14, respectively
Summary
C virus (HCV) infects more than 71 million people worldwide [1], which can lead to liver failure and hepatocellular carcinoma and presents a major global health burden. HCV was discovered in humans 20 years ago [4] but for a long time, researchers failed to identify an animal viral homologue. E subsequently discovered RHV (RHV-rn1) from Norway rats was used to make a small animal model for hepaciviral infection, utilising a reverse genetics approach. In this system, the researchers showed its hepatotropic replication in inbred and outbred rat strains [15]. Emulating HCV infection, they showed that persistent infection leads to gradual liver damage and that the HCV antiviral drug sofosbuvir suppresses replication of RHV-rn. Emulating HCV infection, they showed that persistent infection leads to gradual liver damage and that the HCV antiviral drug sofosbuvir suppresses replication of RHV-rn1. is model can be used to study the mechanisms of HCV persistence, immunity, and pathogenesis
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
