{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T18:49:02Z","timestamp":1776365342668,"version":"3.51.2"},"update-to":[{"DOI":"10.1371\/journal.ppat.1014127","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T00:00:00Z","timestamp":1776297600000}}],"reference-count":44,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T00:00:00Z","timestamp":1775606400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100020963","name":"Moonshot Research and Development Program","doi-asserted-by":"publisher","award":["JPMJMS2025"],"award-info":[{"award-number":["JPMJMS2025"]}],"id":[{"id":"10.13039\/501100020963","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["24fk0210109"],"award-info":[{"award-number":["24fk0210109"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["25fk021016"],"award-info":[{"award-number":["25fk021016"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP25K02498"],"award-info":[{"award-number":["JP25K02498"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP25K10374"],"award-info":[{"award-number":["JP25K10374"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012030","name":"Yakult Honsha","doi-asserted-by":"publisher","award":["Scholarship donation 2025"],"award-info":[{"award-number":["Scholarship donation 2025"]}],"id":[{"id":"10.13039\/501100012030","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    The lack of immuno-competent animal models of hepatitis C virus (HCV) infection has been an obstacle to vaccine development and research on immune responses.\n                    <jats:italic>Hepacivirus ratti<\/jats:italic>\n                    (Norway rat hepacivirus-1: NRHV1) is a virus closely related to HCV that specifically infects the liver and induces hepatocellular carcinoma in rats, making it a promising surrogate model for HCV. NRHV1 expressing a reporter gene serves as a powerful tool for analyzing the\n                    <jats:italic>in vivo<\/jats:italic>\n                    dynamics and pathogenicity mechanisms of NRHV1. In this study, we developed a reporter NRHV1 capable of infection and replication in both immunodeficient mice and cultured cells and established a platform for generating diverse reporter viruses. A reporter virus containing the HiBiT gene in the coding region of NS5A domain III was constructed using circular polymerase extension reaction (CPER). Infection with this reporter virus led to HiBiT activity in infected cells and the activity was correlated with the amount of intracellular viral RNA. In addition, this reporter virus established persistent infection in NOD-SCID mice and led to the generation of HiBiT activity in the livers of infected mice, although loss of the HiBiT gene was observed in some mice. Furthermore, reporter virus recovered from infected mice could infect and generate HiBiT activity in cultured cells. These results demonstrate that the reporter virus can infect hepatocytes\n                    <jats:italic>in vivo<\/jats:italic>\n                    and\n                    <jats:italic>in vitro<\/jats:italic>\n                    . This platform provides a versatile tool for\n                    <jats:italic>in vitro<\/jats:italic>\n                    quantitative antiviral screening and for exploring viral infection dynamics\n                    <jats:italic>in vivo<\/jats:italic>\n                    , although further studies will be required to improve the long-term stability of the HiBiT reporter.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1014127","type":"journal-article","created":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T17:48:21Z","timestamp":1775670501000},"page":"e1014127","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Generation of a HiBiT-expressing recombinant rat hepacivirus supporting both in vivo and in vitro infection"],"prefix":"10.1371","volume":"22","author":[{"given":"Yasunori","family":"Akaike","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomohisa","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hirotake","family":"Kasai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsuya","family":"Yamashita","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshiharu","family":"Matsuura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9542-5188","authenticated-orcid":true,"given":"Kohji","family":"Moriishi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,4,8]]},"reference":[{"issue":"12","key":"ppat.1014127.ref001","doi-asserted-by":"crossref","DOI":"10.3390\/v16121899","article-title":"Chronic Hepatitis C Infection Treated with Direct-Acting Antiviral Agents and Occurrence\/Recurrence of Hepatocellular Carcinoma: Does It Still Matter?","volume":"16","author":"C Smirne","year":"2024","journal-title":"Viruses"},{"issue":"8","key":"ppat.1014127.ref002","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1038\/s41575-022-00608-8","article-title":"Breakthroughs in hepatitis C research: from discovery to cure","volume":"19","author":"MP Manns","year":"2022","journal-title":"Nat Rev Gastroenterol Hepatol"},{"issue":"6","key":"ppat.1014127.ref003","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.3390\/cancers12061351","article-title":"Risk of Hepatocellular Carcinoma after HCV Clearance by Direct-Acting Antivirals Treatment Predictive Factors and Role of Epigenetics","volume":"12","author":"L Rinaldi","year":"2020","journal-title":"Cancers (Basel)"},{"issue":"11","key":"ppat.1014127.ref004","doi-asserted-by":"crossref","first-page":"3869","DOI":"10.3390\/ijms21113869","article-title":"Animal Models Used in Hepatitis C Virus Research","volume":"21","author":"KA Berggren","year":"2020","journal-title":"Int J Mol Sci"},{"issue":"6","key":"ppat.1014127.ref005","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1002\/hep.20268","article-title":"A critical role for the chimpanzee model in the study of hepatitis C","volume":"39","author":"J Bukh","year":"2004","journal-title":"Hepatology"},{"issue":"2","key":"ppat.1014127.ref006","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1006\/viro.1998.9127","article-title":"Transmission of hepatitis C virus infection to tree shrews","volume":"244","author":"ZC Xie","year":"1998","journal-title":"Virology"},{"issue":"7466","key":"ppat.1014127.ref007","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1038\/nature12427","article-title":"Completion of the entire 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Firth","year":"2014","journal-title":"mBio"},{"issue":"2","key":"ppat.1014127.ref011","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1002\/hep.29494","article-title":"Viral persistence, liver disease, and host response in a hepatitis C-like virus rat model","volume":"68","author":"S Trivedi","year":"2018","journal-title":"Hepatology"},{"issue":"6347","key":"ppat.1014127.ref012","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1126\/science.aal1962","article-title":"Mouse models of acute and chronic hepacivirus infection","volume":"357","author":"E Billerbeck","year":"2017","journal-title":"Science"},{"issue":"4","key":"ppat.1014127.ref013","doi-asserted-by":"crossref","first-page":"2367","DOI":"10.1128\/JVI.03162-12","article-title":"A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity","volume":"87","author":"J 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Interactions with Scavenger Receptor Class B Type I","volume":"90","author":"A Zuiani","year":"2016","journal-title":"J Virol"},{"issue":"30","key":"ppat.1014127.ref029","doi-asserted-by":"crossref","first-page":"22725","DOI":"10.1074\/jbc.R110.125294","article-title":"Hepatitis C virus non-structural protein 3 (HCV NS3): a multifunctional antiviral target","volume":"285","author":"KD Raney","year":"2010","journal-title":"J Biol Chem"},{"issue":"2","key":"ppat.1014127.ref030","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S0092-8674(00)81351-3","article-title":"Crystal structure of the hepatitis C virus NS3 protease domain complexed with a synthetic NS4A cofactor peptide","volume":"87","author":"JL Kim","year":"1996","journal-title":"Cell"},{"issue":"1","key":"ppat.1014127.ref031","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.antiviral.2010.11.004","article-title":"Measuring antiviral activity of benzimidazole molecules that alter IRES RNA structure with an infectious hepatitis C virus chimera expressing Renilla luciferase","volume":"89","author":"S Liu","year":"2011","journal-title":"Antiviral Res"},{"key":"ppat.1014127.ref032","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1099\/vir.0.055772-0","article-title":"Direct visualization of hepatitis C virus-infected Huh7.5 cells with a high titre of infectious chimeric JFH1-EGFP reporter virus in three-dimensional Matrigel cell cultures","volume":"95","author":"S Liu","year":"2014","journal-title":"J Gen Virol"},{"key":"ppat.1014127.ref033","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.virol.2017.04.003","article-title":"Sensitive luminescent reporter viruses reveal appreciable release of hepatitis C virus NS5A protein into the extracellular environment","volume":"507","author":"NS 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