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dc.contributor.authorRink, Marco R
dc.contributor.authorBaptista, Marisa A P
dc.contributor.authorFlomm, Felix J
dc.contributor.authorHennig, Thomas
dc.contributor.authorWhisnant, Adam W
dc.contributor.authorWolf, Natalia
dc.contributor.authorSeibel, Jürgen
dc.contributor.authorDölken, Lars
dc.contributor.authorBosse, Jens B
dc.date.accessioned2021-10-01T13:02:56Z
dc.date.available2021-10-01T13:02:56Z
dc.date.issued2021-08-04
dc.identifier.citation. PLoS One. 2021 Aug 4;16(8):e0244166. doi: 10.1371/journal.pone.0244166.en_US
dc.identifier.pmid34347781
dc.identifier.doi10.1371/journal.pone.0244166
dc.identifier.urihttp://hdl.handle.net/10033/623056
dc.description.abstractFluorogenic aptamers are an alternative to established methodology for real-time imaging of RNA transport and dynamics. We developed Broccoli-aptamer concatemers ranging from 4 to 128 substrate-binding site repeats and characterized their behavior fused to an mCherry-coding mRNA in transient transfection, stable expression, and in recombinant cytomegalovirus infection. Concatemerization of substrate-binding sites increased Broccoli fluorescence up to a concatemer length of 16 copies, upon which fluorescence did not increase and mCherry signals declined. This was due to the combined effects of RNA aptamer aggregation and reduced RNA stability. Unfortunately, both cellular and cytomegalovirus genomes were unable to maintain and express high Broccoli concatemer copy numbers, possibly due to recombination events. Interestingly, negative effects of Broccoli concatemers could be partially rescued by introducing linker sequences in between Broccoli repeats warranting further studies. Finally, we show that even though substrate-bound Broccoli is easily photobleached, it can still be utilized in live-cell imaging by adapting a time-lapse imaging protocol.en_US
dc.language.isoenen_US
dc.publisherPLOSen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.titleConcatemeric Broccoli reduces mRNA stability and induces aggregates.en_US
dc.typeArticleen_US
dc.identifier.eissn1932-6203
dc.contributor.departmentHIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.en_US
dc.identifier.journalPloS oneen_US
dc.source.volume16
dc.source.issue8
dc.source.beginpagee0244166
dc.source.endpage
refterms.dateFOA2021-10-01T13:02:56Z
dc.source.journaltitlePloS one
dc.source.countryUnited States


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