A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Detection of Viral −RNA and +RNA Strands in Enterovirus-Infected Cells and Tissues (2020)

Salmikangas, S., Laiho, J. E., Kalander, K., Laajala, M., Honkimaa, A., Shanina, I., Oikarinen, S., Horwitz, M. S., Hyöty, H., & Marjomäki, V. (2020). Detection of Viral −RNA and +RNA Strands in Enterovirus-Infected Cells and Tissues. Microorganisms, 8(12), Article 1928. https://doi.org/10.3390/microorganisms8121928

JYU-tekijät tai -toimittajat

Julkaisun tiedot

Julkaisun kaikki tekijät tai toimittajat: Salmikangas, Sami; Laiho, Jutta E.; Kalander, Kerttu; Laajala, Mira; Honkimaa, Anni; Shanina, Iryna; Oikarinen, Sami; Horwitz, Marc S.; Hyöty, Heikki; Marjomäki, Varpu

Lehti tai sarja: Microorganisms

eISSN: 2076-2607

Julkaisuvuosi: 2020

Ilmestymispäivä: 04.12.2020

Volyymi: 8

Lehden numero: 12

Artikkelinumero: 1928

Kustantaja: MDPI AG

Julkaisumaa: Sveitsi

Julkaisun kieli: englanti

DOI: https://doi.org/10.3390/microorganisms8121928

Julkaisun avoin saatavuus: Avoimesti saatavilla

Julkaisukanavan avoin saatavuus: Kokonaan avoin julkaisukanava

Julkaisu on rinnakkaistallennettu (JYX): https://jyx.jyu.fi/handle/123456789/73337

Julkaisu on rinnakkaistallennettu: http://hdl.handle.net/10138/322686

Lisätietoja: Supplementary Materials:The following are available online at http://www.mdpi.com/2076-2607/8/12/1928/s1:Table S1: Antibodies used in the immunofluorescence and bDNA FISH. Table S2: ViewRNA probe set sequences.Table S3: Components of the RT reaction. Table S4: Components of the qPCR reaction. Figure S1: Cell viabilityassay for antiviral drugs Cal-I1 and Rac1-I. Figure S2: Concentration series of RNA isolated from purified CVA9and detected by qPCR

Tiivistelmä

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 −RNA and +RNA during acute and persistent enterovirus infections. According to our results, the replication of the viral RNA started 2–3 h after infection and the translation shortly after at 3–4 h post-infection. The replication hotspots with newly emerging −RNA 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 −RNA and +RNA strands was almost identical, and −RNA 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.

YSO-asiasanat: enterovirukset; infektiot; RNA; replikaatio

Vapaat asiasanat: antiviral drugs; branched DNA; enterovirus; in situ hybridization; negative RNA; positive RNA; replication

Liittyvät organisaatiot

OKM-raportointi: Kyllä

Raportointivuosi: 2020

JUFO-taso: 1