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dc.contributor.authorRand, Ulfert
dc.contributor.authorKubsch, Tobias
dc.contributor.authorKasmapour, Bahram
dc.contributor.authorCicin-Sain, Luka
dc.date.accessioned2021-02-08T14:49:40Z
dc.date.available2021-02-08T14:49:40Z
dc.date.issued2021-01-08
dc.identifier.citationFront Cell Infect Microbiol. 2021 Jan 8;10:536150. doi: 10.3389/fcimb.2020.536150.en_US
dc.identifier.pmid33489928
dc.identifier.doi10.3389/fcimb.2020.536150
dc.identifier.urihttp://hdl.handle.net/10033/622724
dc.description.abstractHuman Cytomegalovirus (HCMV) infection may result in severe outcomes in immunocompromised individuals such as AIDS patients, transplant recipients, and neonates. To date, no vaccines are available and there are only few drugs for anti-HCMV therapy. Adverse effects and the continuous emergence of drug-resistance strains require the identification of new drug candidates in the near future. Identification and characterization of such compounds and biological factors requires sensitive and reliable detection techniques of HCMV infection, gene expression and spread. In this work, we present and validate a novel concept for multi-reporter herpesviruses, identified through iterative testing of minimally invasive mutations. We integrated up to three fluorescence reporter genes into replication-competent HCMV strains, generating reporter HCMVs that allow the visualization of replication cycle stages of HCMV, namely the immediate early (IE), early (E), and late (L) phase. Fluorescent proteins with clearly distinguishable emission spectra were linked by 2A peptides to essential viral genes, allowing bicistronic expression of the viral and the fluorescent protein without major effects on viral fitness. By using this triple color reporter HCMV, we monitored gene expression dynamics of the IE, E, and L genes by measuring the fluorescent signal of the viral gene-associated fluorophores within infected cell populations and at high temporal resolution. We demonstrate distinct inhibitory profiles of foscarnet, fomivirsen, phosphonoacetic acid, ganciclovir, and letermovir reflecting their mode-of-action. In conclusion, our data argues that this experimental approach allows the identification and characterization of new drug candidates in a single step.en_US
dc.language.isoenen_US
dc.publisherFrontiersen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectHuman Cytomegalovirusen_US
dc.subjectantiviralsen_US
dc.subjectherpesvirusen_US
dc.subjectin vitro drug testingen_US
dc.subjectletermoviren_US
dc.subjectlive-cell imagingen_US
dc.subjectreporter assayen_US
dc.subjectscreeningen_US
dc.titleA Novel Triple-Fluorescent HCMV Strain Reveals Gene Expression Dynamics and Anti-Herpesviral Drug Mechanisms.en_US
dc.typeArticleen_US
dc.identifier.eissn2235-2988
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalFrontiers in cellular and infection microbiologyen_US
dc.source.volume10
dc.source.beginpage536150
dc.source.endpage
refterms.dateFOA2021-02-08T14:49:41Z
dc.source.journaltitleFrontiers in cellular and infection microbiology
dc.source.countrySwitzerland


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Attribution 4.0 International
Except where otherwise noted, this item's license is described as Attribution 4.0 International