{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T14:48:43Z","timestamp":1787496523008,"version":"build-2736575974"},"reference-count":54,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,7,24]],"date-time":"2017-07-24T00:00:00Z","timestamp":1500854400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,7,24]],"date-time":"2017-07-24T00:00:00Z","timestamp":1500854400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    We evaluated Sofosbuvir (SOF), the anti-hepatitis C virus prodrug of \u03b2-d-2\u2032-deoxy-2\u2032-\u03b1-fluoro-2\u2032-\u03b2-C-methyluridine-5\u2032-monophosphate, for potential inhibitory activity against DENV replication. Both cell-based and biochemical assays, based on use of purified DENV full-length NS5 enzyme, were studied. Cytopathic effect protection and virus yield reduction assays confirmed that SOF possessed anti-DENV activity in cell culture with a 50% effective concentration (EC\n                    <jats:sub>50<\/jats:sub>\n                    ) of 4.9\u2009\u00b5M and 1.4\u2009\u00b5M respectively. Real-time RT-PCR verified that SOF inhibits generation of viral RNA with an EC\n                    <jats:sub>50<\/jats:sub>\n                    of 9.9\u2009\u00b5M. Purified DENV NS5 incorporated the active triphosphate form (SOF-TP) into nascent RNA, causing chain-termination. Relative to the natural UTP, the incorporation efficiency of SOF-TP was low (discrimination value\u2009=\u2009327.5). In a primer extension assay, SOF-TP was active against DENV NS5 wild-type polymerase activity with an IC\n                    <jats:sub>50<\/jats:sub>\n                    of 14.7\u2009\u00b1\u20092.5\u2009\u00b5M. The S600T substitution in the B Motif of DENV polymerase conferred 4.3-fold resistance to SOF-TP; this was due to decreased incorporation efficiency rather than enhanced excision of the incorporated SOF nucleotide. SOF has antiviral activity against DENV replication. The high discrimination value in favor of UTP in enzyme assays may not necessarily preclude antiviral activity in cells. SOF may be worthy of evaluation against severe DENV infections in humans.\n                  <\/jats:p>","DOI":"10.1038\/s41598-017-06612-2","type":"journal-article","created":{"date-parts":[[2017,7,18]],"date-time":"2017-07-18T12:07:17Z","timestamp":1500379637000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":68,"title":["Evaluation of Sofosbuvir (\u03b2-D-2\u2032-deoxy-2\u2032-\u03b1-fluoro-2\u2032-\u03b2-C-methyluridine) as an inhibitor of Dengue virus replication\n                #"],"prefix":"10.1038","volume":"7","author":[{"given":"Hong-Tao","family":"Xu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan P.","family":"Colby-Germinario","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Said A.","family":"Hassounah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clare","family":"Fogarty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan","family":"Osman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0369-2316","authenticated-orcid":false,"given":"Navaneethan","family":"Palanisamy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingshan","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maureen","family":"Oliveira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yudong","family":"Quan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark A.","family":"Wainberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,7,24]]},"reference":[{"key":"6612_CR1","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1016\/S0140-6736(14)60572-9","volume":"385","author":"MG Guzman","year":"2015","unstructured":"Guzman, M. G. & Harris, E. Dengue. Lancet \n                           385, 453\u2013465, doi:10.1016\/S0140-6736(14)60572-9 (2015).","journal-title":"Lancet"},{"key":"6612_CR2","doi-asserted-by":"publisher","first-page":"504","DOI":"10.1038\/nature12060","volume":"496","author":"S Bhatt","year":"2013","unstructured":"Bhatt, S. et al. The global distribution and burden of dengue. Nature \n                           496, 504\u2013507, doi:10.1038\/nature12060 (2013).","journal-title":"Nature"},{"key":"6612_CR3","doi-asserted-by":"crossref","unstructured":"Halstead, S. B. & Cohen, S. N. Dengue Hemorrhagic Fever at 60 Years: Early Evolution of Concepts of Causation and Treatment. Microbiol Mol Biol Rev \n                           79, 281\u2013291, doi:79\/3\/281 (2015).","DOI":"10.1128\/MMBR.00009-15"},{"key":"6612_CR4","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1159\/000149642","volume":"24","author":"EG Westaway","year":"1985","unstructured":"Westaway, E. G. et al. Flaviviridae. Intervirology \n                           24, 183\u2013192 (1985).","journal-title":"Intervirology"},{"key":"6612_CR5","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1038\/nrmicro2675","volume":"10","author":"E Decroly","year":"2011","unstructured":"Decroly, E., Ferron, F., Lescar, J. & Canard, B. Conventional and unconventional mechanisms for capping viral mRNA. Nat Rev Microbiol \n                           10, 51\u201365, doi:10.1038\/nrmicro2675 (2011).","journal-title":"Nat Rev Microbiol"},{"key":"6612_CR6","doi-asserted-by":"publisher","first-page":"39926","DOI":"10.1074\/jbc.M104248200","volume":"276","author":"M Ackermann","year":"2001","unstructured":"Ackermann, M. & Padmanabhan, R. De novo synthesis of RNA by the dengue virus RNA-dependent RNA polymerase exhibits temperature dependence at the initiation but not elongation phase. J Biol Chem \n                           276, 39926\u201339937, doi:10.1074\/jbc.M104248200 (2001).","journal-title":"J Biol Chem"},{"key":"6612_CR7","doi-asserted-by":"crossref","unstructured":"Kao, C. C., Del Vecchio, A. M. & Zhong, W. De novo initiation of RNA synthesis by a recombinant flaviviridae RNA-dependent RNA polymerase. Virology \n                           253, 1\u20137, doi:S0042682298995174 (1999).","DOI":"10.1006\/viro.1998.9517"},{"key":"6612_CR8","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.antiviral.2008.06.007","volume":"80","author":"H Malet","year":"2008","unstructured":"Malet, H. et al. The flavivirus polymerase as a target for drug discovery. Antiviral Res \n                           80, 23\u201335, doi:10.1016\/j.antiviral.2008.06.007 (2008).","journal-title":"Antiviral Res"},{"key":"6612_CR9","doi-asserted-by":"crossref","unstructured":"Lim, S. P., Noble, C. G. & Shi, P. Y. The dengue virus NS5 protein as a target for drug discovery. Antiviral Res \n                           119, 57\u201367, doi:S0166-3542(15)00090-X (2015).","DOI":"10.1016\/j.antiviral.2015.04.010"},{"key":"6612_CR10","doi-asserted-by":"publisher","first-page":"e1004682","DOI":"10.1371\/journal.ppat.1004682","volume":"11","author":"Y Zhao","year":"2015","unstructured":"Zhao, Y. et al. A crystal structure of the Dengue virus NS5 protein reveals a novel inter-domain interface essential for protein flexibility and virus replication. PLoS Pathog \n                           11, e1004682, doi:10.1371\/journal.ppat.1004682 (2015).","journal-title":"PLoS Pathog"},{"key":"6612_CR11","doi-asserted-by":"publisher","first-page":"31105","DOI":"10.1074\/jbc.M113.508606","volume":"288","author":"SP Lim","year":"2013","unstructured":"Lim, S. P. et al. A crystal structure of the dengue virus non-structural protein 5 (NS5) polymerase delineates interdomain amino acid residues that enhance its thermostability and de novo initiation activities. J Biol Chem \n                           288, 31105\u201331114, doi:10.1074\/jbc.M113.508606 (2013).","journal-title":"J Biol Chem"},{"key":"6612_CR12","doi-asserted-by":"crossref","unstructured":"Yap, T. L. et al. Crystal structure of the dengue virus RNA-dependent RNA polymerase catalytic domain at 1.85-angstrom resolution. J Virol \n                           81, 4753\u20134765, doi:JVI.02283-06 (2007).","DOI":"10.1128\/JVI.02283-06"},{"key":"6612_CR13","doi-asserted-by":"publisher","first-page":"6498","DOI":"10.1073\/pnas.90.14.6498","volume":"90","author":"TA Steitz","year":"1993","unstructured":"Steitz, T. A. & Steitz, J. A. A general two-metal-ion mechanism for catalytic RNA. Proc Natl Acad Sci U S A \n                           90, 6498\u20136502 (1993).","journal-title":"Proc Natl Acad Sci U S A"},{"key":"6612_CR14","doi-asserted-by":"publisher","first-page":"1171","DOI":"10.1517\/13543776.2014.967212","volume":"24","author":"H Beesetti","year":"2014","unstructured":"Beesetti, H., Khanna, N. & Swaminathan, S. Drugs for dengue: a patent review (2010\u20132014). Expert Opin Ther Pat \n                           24, 1171\u20131184, doi:10.1517\/13543776.2014.967212 (2014).","journal-title":"Expert Opin Ther Pat"},{"key":"6612_CR15","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1016\/j.antiviral.2013.09.013","volume":"100","author":"SP Lim","year":"2013","unstructured":"Lim, S. P. et al. Ten years of dengue drug discovery: progress and prospects. Antiviral Res \n                           100, 500\u2013519, doi:10.1016\/j.antiviral.2013.09.013 (2013).","journal-title":"Antiviral Res"},{"key":"6612_CR16","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.antiviral.2010.05.003","volume":"87","author":"DR Latour","year":"2010","unstructured":"Latour, D. R. et al. Biochemical characterization of the inhibition of the dengue virus RNA polymerase by beta-d-2\u2032-ethynyl-7-deaza-adenosine triphosphate. Antiviral Res \n                           87, 213\u2013222, doi:10.1016\/j.antiviral.2010.05.003 (2010).","journal-title":"Antiviral Res"},{"key":"6612_CR17","doi-asserted-by":"publisher","first-page":"20435","DOI":"10.1073\/pnas.0907010106","volume":"106","author":"Z Yin","year":"2009","unstructured":"Yin, Z. et al. An adenosine nucleoside inhibitor of dengue virus. Proc Natl Acad Sci USA \n                           106, 20435\u201320439, doi:10.1073\/pnas.0907010106 (2009).","journal-title":"Proc Natl Acad Sci USA"},{"key":"6612_CR18","doi-asserted-by":"publisher","first-page":"e1005737","DOI":"10.1371\/journal.ppat.1005737","volume":"12","author":"SP Lim","year":"2016","unstructured":"Lim, S. P. et al. Potent Allosteric Dengue Virus NS5 Polymerase Inhibitors: Mechanism of Action and Resistance Profiling. PLoS Pathog \n                           12, e1005737, doi:10.1371\/journal.ppat.1005737 (2016).","journal-title":"PLoS Pathog"},{"key":"6612_CR19","doi-asserted-by":"publisher","first-page":"695","DOI":"10.1128\/CMR.00102-15","volume":"29","author":"E De Clercq","year":"2016","unstructured":"De Clercq, E. & Li, G. Approved Antiviral Drugs over the Past 50 Years. Clin Microbiol Rev \n                           29, 695\u2013747, doi:10.1128\/CMR.00102-15 (2016).","journal-title":"Clin Microbiol Rev"},{"key":"6612_CR20","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1038\/nrgastro.2016.60","volume":"13","author":"M Gotte","year":"2016","unstructured":"Gotte, M. & Feld, J. J. Direct-acting antiviral agents for hepatitis C: structural and mechanistic insights. Nat Rev Gastroenterol Hepatol \n                           13, 338\u2013351, doi:10.1038\/nrgastro.2016.60 (2016).","journal-title":"Nat Rev Gastroenterol Hepatol"},{"key":"6612_CR21","doi-asserted-by":"publisher","first-page":"7202","DOI":"10.1021\/jm100863x","volume":"53","author":"MJ Sofia","year":"2010","unstructured":"Sofia, M. J. et al. Discovery of a beta-d-2\u2032-deoxy-2\u2032-alpha-fluoro-2\u2032-beta-C-methyluridine nucleotide prodrug (PSI-7977) for the treatment of hepatitis C virus. J Med Chem \n                           53, 7202\u20137218, doi:10.1021\/jm100863x (2010).","journal-title":"J Med Chem"},{"key":"6612_CR22","doi-asserted-by":"crossref","unstructured":"Fung, A. et al. Efficiency of Incorporation and Chain Termination Determines the Inhibition Potency of 2\u2032-Modified Nucleotide Analogs against Hepatitis C Virus Polymerase. Antimicrob Agents Chemother \n                           58, 3636\u20133645, doi:AAC.02666-14 (2014).","DOI":"10.1128\/AAC.02666-14"},{"key":"6612_CR23","doi-asserted-by":"publisher","first-page":"771","DOI":"10.1126\/science.1259210","volume":"347","author":"TC Appleby","year":"2015","unstructured":"Appleby, T. C. et al. Viral replication. Structural basis for RNA replication by the hepatitis C virus polymerase. Science \n                           347, 771\u2013775, doi:10.1126\/science.1259210347\/6223\/771 (2015).","journal-title":"Science"},{"key":"6612_CR24","doi-asserted-by":"crossref","unstructured":"Ma, H. et al. Characterization of the metabolic activation of hepatitis C virus nucleoside inhibitor beta-D-2\u2032-Deoxy-2\u2032-fluoro-2\u2032-C-methylcytidine (PSI-6130) and identification of a novel active 5\u2032-triphosphate species. J Biol Chem \n                           282, 29812\u201329820, doi:M705274200 (2007).","DOI":"10.1074\/jbc.M705274200"},{"key":"6612_CR25","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.coviro.2014.08.004","volume":"9","author":"J Deval","year":"2014","unstructured":"Deval, J., Symons, J. A. & Beigelman, L. Inhibition of viral RNA polymerases by nucleoside and nucleotide analogs: therapeutic applications against positive-strand RNA viruses beyond hepatitis C virus. Curr Opin Virol \n                           9, 1\u20137, doi:10.1016\/j.coviro.2014.08.004 (2014).","journal-title":"Curr Opin Virol"},{"key":"6612_CR26","doi-asserted-by":"crossref","unstructured":"Bullard-Feibelman, K. M. et al. The FDA-approved drug sofosbuvir inhibits Zika virus infection. Antiviral Res \n                           137, 134\u2013140, doi:S0166-3542(16)30660-X (2017).","DOI":"10.1016\/j.antiviral.2016.11.023"},{"key":"6612_CR27","doi-asserted-by":"crossref","unstructured":"Xu, H. T. et al. Purification of Zika virus RNA-dependent RNA polymerase and its use to identify small-molecule Zika inhibitors. J Antimicrob Chemother, doi:dkw514 (2017).","DOI":"10.1093\/jac\/dkw514"},{"key":"6612_CR28","doi-asserted-by":"publisher","first-page":"40920","DOI":"10.1038\/srep40920","volume":"7","author":"CQ Sacramento","year":"2017","unstructured":"Sacramento, C. Q. et al. The clinically approved antiviral drug sofosbuvir inhibits Zika virus replication. Sci Rep \n                           7, 40920, doi:10.1038\/srep40920 (2017).","journal-title":"Sci Rep"},{"key":"6612_CR29","doi-asserted-by":"publisher","first-page":"600","DOI":"10.1128\/AAC.02203-15","volume":"60","author":"HT Xu","year":"2015","unstructured":"Xu, H. T. et al. Identification of a Pyridoxine-Derived Small-Molecule Inhibitor Targeting Dengue Virus RNA-Dependent RNA Polymerase. Antimicrob Agents Chemother \n                           60, 600\u2013608, doi:10.1128\/AAC.02203-15 (2015).","journal-title":"Antimicrob Agents Chemother"},{"key":"6612_CR30","doi-asserted-by":"crossref","unstructured":"Diamond, M. S., Zachariah, M. & Harris, E. Mycophenolic acid inhibits dengue virus infection by preventing replication of viral RNA. Virology \n                           304, 211\u2013221, doi:S0042682202916855 (2002).","DOI":"10.1006\/viro.2002.1685"},{"key":"6612_CR31","doi-asserted-by":"crossref","unstructured":"Takhampunya, R., Ubol, S., Houng, H. S., Cameron, C. E. & Padmanabhan, R. Inhibition of dengue virus replication by mycophenolic acid and ribavirin. J Gen Virol \n                           87, 1947\u20131952, doi:87\/7\/1947 (2006).","DOI":"10.1099\/vir.0.81655-0"},{"key":"6612_CR32","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1016\/j.antiviral.2015.04.007","volume":"119","author":"P Niyomrattanakit","year":"2015","unstructured":"Niyomrattanakit, P. et al. Stabilization of dengue virus polymerase in de novo initiation assay provides advantages for compound screening. Antiviral Res \n                           119, 36\u201346, doi:10.1016\/j.antiviral.2015.04.007 (2015).","journal-title":"Antiviral Res"},{"key":"6612_CR33","doi-asserted-by":"crossref","unstructured":"Selisko, B. et al. Comparative mechanistic studies of de novo RNA synthesis by flavivirus RNA-dependent RNA polymerases. Virology \n                           351, 145\u2013158, doi:S0042-6822(06)00197-8 (2006).","DOI":"10.1016\/j.virol.2006.03.026"},{"key":"6612_CR34","doi-asserted-by":"publisher","first-page":"11642","DOI":"10.1093\/nar\/gku666","volume":"42","author":"S Potisopon","year":"2014","unstructured":"Potisopon, S. et al. The methyltransferase domain of dengue virus protein NS5 ensures efficient RNA synthesis initiation and elongation by the polymerase domain. Nucleic Acids Res \n                           42, 11642\u201311656, doi:10.1093\/nar\/gku666 (2014).","journal-title":"Nucleic Acids Res"},{"key":"6612_CR35","doi-asserted-by":"crossref","unstructured":"Potisopon, S., Ferron, F., Fattorini, V., Selisko, B. & Canard, B. Substrate selectivity of Dengue and Zika virus NS5 polymerase towards 2\u2032-modified nucleotide analogues. Antiviral Res, doi:S0166-3542(16)30754-9 (2016).","DOI":"10.1016\/j.antiviral.2016.12.021"},{"key":"6612_CR36","doi-asserted-by":"crossref","unstructured":"Lu, G. et al. Analysis of Ribonucleotide 5\u2032 -Triphosphate Analogs as Potential Inhibitors of Zika Virus RNA-dependent RNA Polymerase Using Non-Radioactive Polymerase Assays. Antimicrob Agents Chemother, doi:AAC.01967-16 (2016).","DOI":"10.1128\/AAC.01967-16"},{"key":"6612_CR37","doi-asserted-by":"crossref","unstructured":"Dutartre, H., Bussetta, C., Boretto, J. & Canard, B. General catalytic deficiency of hepatitis C virus RNA polymerase with an S282T mutation and mutually exclusive resistance towards 2\u2032-modified nucleotide analogues. Antimicrob Agents Chemother \n                           50, 4161\u20134169, doi:AAC.00433-06 (2006).","DOI":"10.1128\/AAC.00433-06"},{"key":"6612_CR38","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jmb.2004.01.030","volume":"337","author":"CM D\u2019Abramo","year":"2004","unstructured":"D\u2019Abramo, C. M., Cellai, L. & Gotte, M. Excision of incorporated nucleotide analogue chain-terminators can diminish their inhibitory effects on viral RNA-dependent RNA polymerases. J Mol Biol \n                           337, 1\u201314, doi:10.1016\/j.jmb.2004.01.030 (2004).","journal-title":"J Mol Biol"},{"key":"6612_CR39","doi-asserted-by":"crossref","unstructured":"Deval, J., Powdrill, M. H., D\u2019Abramo, C. M., Cellai, L. & Gotte, M. Pyrophosphorolytic excision of nonobligate chain terminators by hepatitis C virus NS5B polymerase. Antimicrob Agents Chemother \n                           51, 2920\u20132928, doi:AAC.00186-07 (2007).","DOI":"10.1128\/AAC.00186-07"},{"key":"6612_CR40","doi-asserted-by":"publisher","first-page":"E348","DOI":"10.1073\/pnas.1214924110","volume":"110","author":"Z Jin","year":"2013","unstructured":"Jin, Z., Leveque, V., Ma, H., Johnson, K. A. & Klumpp, K. NTP-mediated nucleotide excision activity of hepatitis C virus RNA-dependent RNA polymerase. Proc Natl Acad Sci USA \n                           110, E348\u2013357, doi:10.1073\/pnas.1214924110 (2013).","journal-title":"Proc Natl Acad Sci USA"},{"key":"6612_CR41","doi-asserted-by":"publisher","first-page":"1375","DOI":"10.1038\/82191","volume":"6","author":"S Crotty","year":"2000","unstructured":"Crotty, S. et al. The broad-spectrum antiviral ribonucleoside ribavirin is an RNA virus mutagen. Nat Med \n                           6, 1375\u20131379, doi:10.1038\/82191 (2000).","journal-title":"Nat Med"},{"key":"6612_CR42","first-page":"218","volume":"3","author":"HS Te","year":"2007","unstructured":"Te, H. S., Randall, G. & Jensen, D. M. Mechanism of action of ribavirin in the treatment of chronic hepatitis C. Gastroenterol Hepatol (NY) \n                           3, 218\u2013225 (2007).","journal-title":"Gastroenterol Hepatol (NY)"},{"key":"6612_CR43","doi-asserted-by":"crossref","unstructured":"Fraser, J. E. et al. A Nuclear Transport Inhibitor That Modulates the Unfolded Protein Response and Provides In Vivo Protection Against Lethal Dengue virus Infection. J Infect Dis doi:jiu319 (2014).","DOI":"10.1093\/infdis\/jiu319"},{"issue":"Suppl 5","key":"6612_CR44","doi-asserted-by":"publisher","first-page":"S179","DOI":"10.1016\/j.dld.2014.09.026","volume":"46","author":"A Mangia","year":"2014","unstructured":"Mangia, A. & Piazzolla, V. Overall efficacy and safety results of sofosbuvir-based therapies in phase II and III studies. Dig Liver Dis \n                           46(Suppl 5), S179\u2013185, doi:10.1016\/j.dld.2014.09.026 (2014).","journal-title":"Dig Liver Dis"},{"key":"6612_CR45","doi-asserted-by":"publisher","first-page":"2599","DOI":"10.1056\/NEJMoa1512610","volume":"373","author":"JJ Feld","year":"2015","unstructured":"Feld, J. J. et al. Sofosbuvir and Velpatasvir for HCV Genotype 1, 2, 4, 5, and 6 Infection. N Engl J Med \n                           373, 2599\u20132607, doi:10.1056\/NEJMoa1512610 (2015).","journal-title":"N Engl J Med"},{"key":"6612_CR46","doi-asserted-by":"publisher","unstructured":"Medina, F. et al. Dengue virus: isolation, propagation, quantification, and storage. Curr Protoc Microbiol \n                           Chapter 15, Unit 15D 12, doi:10.1002\/9780471729259.mc15d02s27 (2012).","DOI":"10.1002\/9780471729259.mc15d02s27"},{"key":"6612_CR47","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1016\/j.virusres.2013.12.001","volume":"180","author":"JA Usme-Ciro","year":"2014","unstructured":"Usme-Ciro, J. A., Lopera, J. A., Enjuanes, L., Almazan, F. & Gallego-Gomez, J. C. Development of a novel DNA-launched dengue virus type 2 infectious clone assembled in a bacterial artificial chromosome. Virus Res \n                           180, 12\u201322, doi:10.1016\/j.virusres.2013.12.001 (2014).","journal-title":"Virus Res"},{"key":"6612_CR48","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1007\/978-1-62703-484-5_19","volume":"1030","author":"EY Gong","year":"2013","unstructured":"Gong, E. Y. et al. Expression and purification of dengue virus NS5 polymerase and development of a high-throughput enzymatic assay for screening inhibitors of dengue polymerase. Methods Mol Biol \n                           1030, 237\u2013247, doi:10.1007\/978-1-62703-484-5_19 (2013).","journal-title":"Methods Mol Biol"},{"key":"6612_CR49","doi-asserted-by":"publisher","first-page":"45389","DOI":"10.1074\/jbc.M405072200","volume":"279","author":"PR Meyer","year":"2004","unstructured":"Meyer, P. R., Smith, A. J., Matsuura, S. E. & Scott, W. A. Effects of primer-template sequence on ATP-dependent removal of chain-terminating nucleotide analogues by HIV-1 reverse transcriptase. J Biol Chem \n                           279, 45389\u201345398, doi:10.1074\/jbc.M405072200 (2004).","journal-title":"J Biol Chem"},{"key":"6612_CR50","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1186\/1471-2105-9-40","volume":"9","author":"Y Zhang","year":"2008","unstructured":"Zhang, Y. I-TASSER server for protein 3D structure prediction. BMC Bioinformatics \n                           9, 40, doi:10.1186\/1471-2105-9-40 (2008).","journal-title":"BMC Bioinformatics"},{"key":"6612_CR51","doi-asserted-by":"publisher","first-page":"2588","DOI":"10.1093\/bioinformatics\/btt447","volume":"29","author":"J Yang","year":"2013","unstructured":"Yang, J., Roy, A. & Zhang, Y. Protein-ligand binding site recognition using complementary binding-specific substructure comparison and sequence profile alignment. Bioinformatics \n                           29, 2588\u20132595, doi:10.1093\/bioinformatics\/btt447 (2013).","journal-title":"Bioinformatics"},{"key":"6612_CR52","doi-asserted-by":"publisher","first-page":"1605","DOI":"10.1002\/jcc.20084","volume":"25","author":"EF Pettersen","year":"2004","unstructured":"Pettersen, E. F. et al. UCSF Chimera\u2013a visualization system for exploratory research and analysis. J Comput Chem \n                           25, 1605\u20131612, doi:10.1002\/jcc.20084 (2004).","journal-title":"J Comput Chem"},{"key":"6612_CR53","doi-asserted-by":"publisher","first-page":"455","DOI":"10.1002\/jcc.21334","volume":"31","author":"O Trott","year":"2010","unstructured":"Trott, O. & Olson, A. J. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem \n                           31, 455\u2013461, doi:10.1002\/jcc.21334 (2010).","journal-title":"J Comput Chem"},{"key":"6612_CR54","doi-asserted-by":"publisher","first-page":"1548","DOI":"10.1093\/bioinformatics\/18.11.1548","volume":"18","author":"SS Sheik","year":"2002","unstructured":"Sheik, S. S., Sundararajan, P., Hussain, A. S. & Sekar, K. Ramachandran plot on the web. Bioinformatics \n                           18, 1548\u20131549 (2002).","journal-title":"Bioinformatics"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-06612-2","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-06612-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-06612-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T02:24:10Z","timestamp":1671762250000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-06612-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,7,24]]},"references-count":54,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["6612"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-06612-2","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,7,24]]},"assertion":[{"value":"10 March 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 May 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 July 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"6345"}}