{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T21:31:17Z","timestamp":1770413477761,"version":"3.49.0"},"reference-count":25,"publisher":"Proceedings of the National Academy of Sciences","issue":"12","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,3,22]]},"abstract":"<jats:p>MicroRNA-122 (miR-122) is believed to stimulate hepatitis C virus (HCV) replication through interaction with two adjacent sites downstream of stem loop I (SLI) within the HCV 5\u2032 untranslated region (5\u2032 UTR). Recently, it was demonstrated that locked nucleic acid SPC3649-induced miR-122 antagonism suppressed HCV genotype 1a and 1b infection in vivo. However, virus-producing culture systems with 5\u2032 UTR of different HCV genotypes have not been available for testing 5\u2032 UTR-based treatment approaches. Using JFH1-based Core-NS2 genotype recombinants, we developed 5\u2032 UTR-NS2 recombinants of HCV genotypes 1a, 1b, 2a, 2b, 3a, 4a, 5a, and 6a with efficient growth in Huh7.5 cells. Deletion mutagenesis studies demonstrated that the 5\u2032 UTR SLI was essential for genotypes 1\u20136 infection. However, lack of SLI could be compensated for by insertion of other structured HCV or host RNA sequences, including U3 small nucleolar RNA. We demonstrated that SPC3649-induced miR-122 antagonism had a potent antiviral effect against HCV genotypes 1\u20136 5\u2032 UTR-NS2 viruses. Strikingly, HCV recombinant virus with substitution of SLI and miR-122 binding site 1 (S1) by the U3 RNA sequence was not affected by miR-122 antagonism; this was attributed to the lack of an intact S1 by reverse genetics studies. Therefore, we engineered the corresponding U3 RNA sequences into S1 and demonstrated that HCV recombinants with wild-type SLI and single or combined mutations at four of eight nucleotides of S1 were viable in Huh7.5 cells. These mutations reduced the efficacy of SPC3649 treatment, indicating that escape variants to miR-122 antagonism-based HCV therapy could potentially occur.<\/jats:p>","DOI":"10.1073\/pnas.1016606108","type":"journal-article","created":{"date-parts":[[2011,3,8]],"date-time":"2011-03-08T12:01:21Z","timestamp":1299585681000},"page":"4991-4996","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":160,"title":["MicroRNA-122 antagonism against hepatitis C virus genotypes 1\u20136 and reduced efficacy by host RNA insertion or mutations in the HCV 5\u2032 UTR"],"prefix":"10.1073","volume":"108","author":[{"given":"Yi-Ping","family":"Li","sequence":"first","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, Hvidovre, and Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2650 Hvidovre, Denmark"}]},{"given":"Judith M.","family":"Gottwein","sequence":"additional","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, Hvidovre, and Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2650 Hvidovre, Denmark"}]},{"given":"Troels K.","family":"Scheel","sequence":"additional","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, Hvidovre, and Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2650 Hvidovre, Denmark"}]},{"given":"Tanja B.","family":"Jensen","sequence":"additional","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, Hvidovre, and Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2650 Hvidovre, Denmark"}]},{"given":"Jens","family":"Bukh","sequence":"additional","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, Hvidovre, and Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2650 Hvidovre, Denmark"}]}],"member":"341","published-online":{"date-parts":[[2011,3,7]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/nrmicro1645","article-title":"Replication of hepatitis C virus","volume":"5","author":"Moradpour D","year":"2007","unstructured":"D Moradpour, F Penin, CM Rice, Replication of hepatitis C virus. 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EMBO J 27, 3300\u20133310 (2008).","journal-title":"EMBO J"},{"key":"e_1_3_4_5_2","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1126\/science.1113329","article-title":"Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA","volume":"309","author":"Jopling CL","year":"2005","unstructured":"CL Jopling, M Yi, AM Lancaster, SM Lemon, P Sarnow, Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA. Science 309, 1577\u20131581 (2005).","journal-title":"Science"},{"key":"e_1_3_4_6_2","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.chom.2008.05.013","article-title":"Position-dependent function for a tandem microRNA miR-122-binding site located in the hepatitis C virus RNA genome","volume":"4","author":"Jopling CL","year":"2008","unstructured":"CL Jopling, S Sch\u00fctz, P Sarnow, Position-dependent function for a tandem microRNA miR-122-binding site located in the hepatitis C virus RNA genome. 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