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However, the full-length J6 clone (J6CF), which we previously found to be fully functional in vivo, was replication incompetent in vitro. Through a systematic approach of culturing J6 with minimal JFH1 sequences, we identified three mutations in NS3, NS4A, and NS5B that permitted full-length J6 propagation and adaptation with infectivity titers comparable to JFH1-based systems. The most efficient recombinant, J6cc, had six adaptive mutations and did not accumulate additional changes following viral passage. We demonstrated that HCV NS3\/NS4A protease-, NS5A- and NS5B polymerase-directed drugs respectively inhibited full-length J6 infection dose dependently. Importantly, the three J6-derived mutations enabled culture adaptation of the genetically divergent isolate J8 (genotype 2b), which differed from the J6 nucleotide sequence by 24%. The most efficient recombinant, J8cc, had nine adaptive mutations and was genetically stable after viral passage. The availability of these robust JFH1-independent genotype 2a and 2b culture systems represents an important advance, and the approach used might permit culture development of other isolates, with implications for improved individualized treatments of HCV patients and for development of broadly efficient vaccines.<\/jats:p>","DOI":"10.1073\/pnas.1203829109","type":"journal-article","created":{"date-parts":[[2012,3,31]],"date-time":"2012-03-31T00:38:19Z","timestamp":1333154299000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":58,"title":["Robust full-length hepatitis C virus genotype 2a and 2b infectious cultures using mutations identified by a systematic approach applicable to patient strains"],"prefix":"10.1073","volume":"109","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, DK-2650 Hvidovre, Denmark;"},{"name":"Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santseharay","family":"Ramirez","sequence":"additional","affiliation":[{"name":"Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases and Clinical Research Centre, Copenhagen University Hospital, DK-2650 Hvidovre, Denmark;"},{"name":"Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"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, DK-2650 Hvidovre, Denmark;"},{"name":"Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Troels K. 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