{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T00:04:42Z","timestamp":1778630682165,"version":"3.51.4"},"reference-count":37,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2018,12,9]],"date-time":"2018-12-09T00:00:00Z","timestamp":1544313600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["AI097318"],"award-info":[{"award-number":["AI097318"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"The Development Program (973) of China","award":["2012CB518900"],"award-info":[{"award-number":["2012CB518900"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Zika virus (ZIKV) is a mosquito-borne flavivirus that has emerged as an important human viral pathogen, causing congenital malformation including microcephaly among infants born to mothers infected with the virus during pregnancy. Phylogenetic analysis suggested that ZIKV can be classified into African and Asian lineages. In this study, we have developed a stable plasmid-based reverse genetic system for robust production of both ZIKV prototype African-lineage MR766 and clinical Asian-lineage FSS13025 strains using a tetracycline (Tet)-controlled gene expression vector. Transcription of the full-length ZIKV RNA is under the control of the Tet-responsive Ptight promoter at the 5\u2032 end and an antigenomic ribozyme of hepatitis delta virus at the 3\u2032 end. The transcription of infectious ZIKV RNA genome was efficiently induced by doxycycline. This novel ZIKV reverse genetics system will be valuable for the study of molecular viral pathogenesis of ZIKV and the development of new vaccines against ZIKV infection.<\/jats:p>","DOI":"10.3390\/v10120700","type":"journal-article","created":{"date-parts":[[2018,12,10]],"date-time":"2018-12-10T03:36:41Z","timestamp":1544413001000},"page":"700","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Tet-Inducible Production of Infectious Zika Virus from the Full-Length cDNA Clones of African- and Asian-Lineage Strains"],"prefix":"10.3390","volume":"10","author":[{"given":"Lizhou","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Ji","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Peking University Health Science Center School of Basic Medical Sciences, Beijing 100191, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuang","family":"Lyu","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luhua","family":"Qiao","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0354-6020","authenticated-orcid":false,"given":"Guangxiang","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294, USA"},{"name":"Department of Microbiology, Peking University Health Science Center School of Basic Medical Sciences, Beijing 100191, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,12,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/0035-9203(52)90042-4","article-title":"Zika virus I. Isolations and serological specificity","volume":"46","author":"Dick","year":"1952","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jcv.2016.09.012","article-title":"An updated review of Zika virus","volume":"84","author":"Abushouk","year":"2016","journal-title":"J. Clin. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Krauer, F., Riesen, M., Reveiz, L., Oladapo, O.T., Martinez-Vega, R., Porgo, T.V., Haefliger, A., Broutet, N.J., Low, N., and Group WHOZCW (2017). Zika virus Infection as a Cause of Congenital Brain Abnormalities and Guillain-Barre Syndrome: Systematic Review. PLoS Med., 14.","DOI":"10.1371\/journal.pmed.1002203"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1016\/S0140-6736(16)30058-7","article-title":"Guillain-Barre syndrome associated with Zika virus infection","volume":"387","author":"Brasil","year":"2016","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1016\/j.cell.2017.04.008","article-title":"Zika Virus Persistence in the Central Nervous System and Lymph Nodes of Rhesus Monkeys","volume":"169","author":"Aid","year":"2017","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.1056\/NEJMoa1613108","article-title":"Persistence of Zika Virus in Body Fluids\u2014Preliminary Report","volume":"379","author":"Rosenberg","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1887","DOI":"10.3201\/eid2110.151125","article-title":"Zika virus Transmission from French Polynesia to Brazil","volume":"21","author":"Musso","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3091","DOI":"10.1016\/j.celrep.2016.11.070","article-title":"A Mouse Model of Zika Virus Sexual Transmission and Vaginal Viral Replication","volume":"17","author":"Tang","year":"2016","journal-title":"Cell Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1720","DOI":"10.1093\/infdis\/jix204","article-title":"Fetal Growth Restriction Caused by Sexual Transmission of Zika Virus in Mice","volume":"215","author":"Uraki","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.chom.2017.01.004","article-title":"Zika virus Pathogenesis and Tissue Tropism","volume":"21","author":"Miner","year":"2017","journal-title":"Cell Host Microbe"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.antiviral.2016.03.010","article-title":"Zika virus: History, emergence, biology, and prospects for control","volume":"130","author":"Weaver","year":"2016","journal-title":"Antivir. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Bowen, J.R., Quicke, K.M., Maddur, M.S., O\u2019Neal, J.T., McDonald, C.E., Fedorova, N.B., Puri, V., Shabman, R.S., Pulendran, B., and Suthar, M.S. (2017). Zika Virus Antagonizes Type I Interferon Responses during Infection of Human Dendritic Cells. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006164"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1410","DOI":"10.4269\/ajtmh.17-0263","article-title":"Differential Neurovirulence of African and Asian Genotype Zika virus Isolates in Outbred Immunocompetent Mice","volume":"97","author":"Duggal","year":"2017","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1038\/nature20556","article-title":"Zika virus infection damages the testes in mice","volume":"540","author":"Govero","year":"2016","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Simonin, Y., van Riel, D., Van de Perre, P., Rockx, B., and Salinas, S. (2017). Differential virulence between Asian and African lineages of Zika virus. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005821"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Schwarz, M.C., Sourisseau, M., Espino, M.M., Gray, E.S., Chambers, M.T., Tortorella, D., and Evans, M.J. (2016). Rescue of the 1947 Zika Virus Prototype Strain with a Cytomegalovirus Promoter-Driven cDNA Clone. mSphere, 1.","DOI":"10.1128\/mSphere.00246-16"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1038\/nm.4322","article-title":"A live-attenuated Zika virus vaccine candidate induces sterilizing immunity in mouse models","volume":"23","author":"Shan","year":"2017","journal-title":"Nat. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1016\/j.chom.2016.05.004","article-title":"An Infectious cDNA Clone of Zika Virus to Study Viral Virulence, Mosquito Transmission, and Antiviral Inhibitors","volume":"19","author":"Shan","year":"2016","journal-title":"Cell Host Microbe"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Tsetsarkin, K.A., Kenney, H., Chen, R., Liu, G., Manukyan, H., Whitehead, S.S., Laassri, M., Chumakov, K., and Pletnev, A.G. (2016). A Full-Length Infectious cDNA Clone of Zika Virus from the 2015 Epidemic in Brazil as a Genetic Platform for Studies of Virus-Host Interactions and Vaccine Development. mBio, 7.","DOI":"10.1128\/mBio.01114-16"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Weger-Lucarelli, J., Duggal, N.K., Bullard-Feibelman, K., Veselinovic, M., Romo, H., Nguyen, C., Ruckert, C., Brault, A.C., Bowen, R.A., and Stenglein, M. (2017). Development and Characterization of Recombinant Virus Generated from a New World Zika Virus Infectious Clone. J. Virol., 91.","DOI":"10.1128\/JVI.00172-17"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Widman, D.G., Young, E., Yount, B.L., Plante, K.S., Gallichotte, E.N., Carbaugh, D.L., Peck, K.M., Plante, J., Swanstrom, J., and Heise, M.T. (2017). A Reverse Genetics Platform That Spans the Zika Virus Family Tree. mBio, 8.","DOI":"10.1128\/mBio.02014-16"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Li, D., Aaskov, J., and Lott, W.B. (2011). Identification of a cryptic prokaryotic promoter within the cDNA encoding the 5\u2032 end of dengue virus RNA genome. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0018197"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Munster, M., Plaszczyca, A., Cortese, M., Neufeldt, C.J., Goellner, S., Long, G., and Bartenschlager, R. (2018). A Reverse Genetics System for Zika Virus Based on a Simple Molecular Cloning Strategy. Viruses, 10.","DOI":"10.3390\/v10070368"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/S0065-3527(08)60348-6","article-title":"Functional cDNA clones of the Flaviviridae: Strategies and applications","volume":"53","author":"Ruggli","year":"1999","journal-title":"Adv. Virus Res."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Liu, Z.Y., Yu, J.Y., Huang, X.Y., Fan, H., Li, X.F., Deng, Y.Q., Ji, X., Cheng, M.L., Ye, Q., and Zhao, H. (2017). Characterization of cis-Acting RNA Elements of Zika Virus by Using a Self-Splicing Ribozyme-Dependent Infectious Clone. J. Virol., 91.","DOI":"10.1128\/JVI.00484-17"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1099\/jgv.0.000862","article-title":"Recovery of the Zika virus through an in vitro ligation approach","volume":"98","author":"Deng","year":"2017","journal-title":"J. Gen. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1766","DOI":"10.1126\/science.7792603","article-title":"Transcriptional activation by tetracyclines in mammalian cells","volume":"268","author":"Gossen","year":"1995","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"13963","DOI":"10.1128\/JVI.79.22.13963-13973.2005","article-title":"Robust production of infectious hepatitis C virus (HCV) from stably HCV cDNA-transfected human hepatoma cells","volume":"79","author":"Cai","year":"2005","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Anfasa, F., Siegers, J.Y., van der Kroeg, M., Mumtaz, N., Stalin Raj, V., de Vrij, F.M.S., Widagdo, W., Gabriel, G., Salinas, S., and Simonin, Y. (2017). Phenotypic Differences between Asian and African Lineage Zika Viruses in Human Neural Progenitor Cells. mSphere, 2.","DOI":"10.1128\/mSphere.00292-17"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Sheridan, M.A., Balaraman, V., Schust, D.J., Ezashi, T., Roberts, R.M., and Franz, A.W.E. (2018). African and Asian strains of Zika virus differ in their ability to infect and lyse primitive human placental trophoblast. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0200086"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.ebiom.2016.09.020","article-title":"Zika Virus Strains Potentially Display Different Infectious Profiles in Human Neural Cells","volume":"12","author":"Simonin","year":"2016","journal-title":"EBioMedicine"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"156","DOI":"10.2174\/1566523216666160524144041","article-title":"Tet-On Systems For Doxycycline-inducible Gene Expression","volume":"16","author":"Das","year":"2016","journal-title":"Curr. Gene Ther."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"De Maio, F.A., Risso, G., Iglesias, N.G., Shah, P., Pozzi, B., Gebhard, L.G., Mammi, P., Mancini, E., Yanovsky, M.J., and Andino, R. (2016). The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005841"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.gene.2017.07.049","article-title":"Molecular cloning and characterization of the genes encoding the proteins of Zika virus","volume":"628","author":"Hou","year":"2017","journal-title":"Gene"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Fulton, B.O., Sachs, D., Schwarz, M.C., Palese, P., and Evans, M.J. (2017). Transposon mutagenesis of the Zika virus genome highlights regions essential for RNA replication and restricted for immune evasion. J. Virol.","DOI":"10.1128\/JVI.00698-17"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.antiviral.2014.12.007","article-title":"Flavivirus reverse genetic systems, construction techniques and applications: A historical perspective","volume":"114","author":"Aubry","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Lewin, A., Mayer, M., Chusainow, J., Jacob, D., and Appel, B. (2005). Viral promoters can initiate expression of toxin genes introduced into Escherichia coli. BMC Biotechnol., 5.","DOI":"10.1186\/1472-6750-5-19"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/12\/700\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:32:24Z","timestamp":1760196744000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/12\/700"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,12,9]]},"references-count":37,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["v10120700"],"URL":"https:\/\/doi.org\/10.3390\/v10120700","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,9]]}}}