{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:28:14Z","timestamp":1760239694573,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2020,12,4]],"date-time":"2020-12-04T00:00:00Z","timestamp":1607040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002341","name":"Academy of Finland","doi-asserted-by":"publisher","award":["288671"],"award-info":[{"award-number":["288671"]}],"id":[{"id":"10.13039\/501100002341","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The current methods to study the distribution and dynamics of viral RNA molecules inside infected cells are not ideal, as electron microscopy and immunohistochemistry can only detect mature virions, and quantitative real-time PCR does not reveal localized distribution of RNAs. We demonstrated here the branched DNA in situ hybridization (bDNA ISH) technology to study both the amount and location of the emerging \u2212RNA and +RNA during acute and persistent enterovirus infections. According to our results, the replication of the viral RNA started 2\u20133 h after infection and the translation shortly after at 3\u20134 h post-infection. The replication hotspots with newly emerging \u2212RNA were located quite centrally in the cell, while the +RNA production and most likely virion assembly took place in the periphery of the cell. We also discovered that the pace of replication of \u2212RNA and +RNA strands was almost identical, and \u2212RNA was absent during antiviral treatments. ViewRNA ISH with our custom probes also showed a good signal during acute and persistent enterovirus infections in cell and mouse models. Considering these results, along with the established bDNA FISH protocol modified by us, the effects of antiviral drugs and the emergence of enterovirus RNAs in general can be studied more effectively.<\/jats:p>","DOI":"10.3390\/microorganisms8121928","type":"journal-article","created":{"date-parts":[[2020,12,4]],"date-time":"2020-12-04T11:59:00Z","timestamp":1607083140000},"page":"1928","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Detection of Viral \u2212RNA and +RNA Strands in Enterovirus-Infected Cells and Tissues"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6264-4587","authenticated-orcid":false,"given":"Sami","family":"Salmikangas","sequence":"first","affiliation":[{"name":"Department of Biological and Environmental Science\/Nanoscience Center, University of Jyv\u00e4skyl\u00e4, Survontie 9C, FI-40500 Jyv\u00e4skyl\u00e4, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5443-4651","authenticated-orcid":false,"given":"Jutta E.","family":"Laiho","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Health Technology, Tampere University, FI-33520 Tampere, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kerttu","family":"Kalander","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Science\/Nanoscience Center, University of Jyv\u00e4skyl\u00e4, Survontie 9C, FI-40500 Jyv\u00e4skyl\u00e4, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3981-280X","authenticated-orcid":false,"given":"Mira","family":"Laajala","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Science\/Nanoscience Center, University of Jyv\u00e4skyl\u00e4, Survontie 9C, FI-40500 Jyv\u00e4skyl\u00e4, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anni","family":"Honkimaa","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Health Technology, Tampere University, FI-33520 Tampere, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iryna","family":"Shanina","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, Vancouver, BC V6T1Z3, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sami","family":"Oikarinen","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Health Technology, Tampere University, FI-33520 Tampere, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc S.","family":"Horwitz","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, Vancouver, BC V6T1Z3, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heikki","family":"Hy\u00f6ty","sequence":"additional","affiliation":[{"name":"Faculty of Medicine and Health Technology, Tampere University, FI-33520 Tampere, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4592-5926","authenticated-orcid":false,"given":"Varpu","family":"Marjom\u00e4ki","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Science\/Nanoscience Center, University of Jyv\u00e4skyl\u00e4, Survontie 9C, FI-40500 Jyv\u00e4skyl\u00e4, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1038\/s41579-018-0005-4","article-title":"The life cycle of non-polio enteroviruses and how to target it","volume":"16","author":"Baggen","year":"2018","journal-title":"Nat. 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