Avoidance of reporter assay distortions from fused dual reporters

dc.contributor.authorLoughran, Gary
dc.contributor.authorHoward, Michael T.
dc.contributor.authorFirth, Andrew E.
dc.contributor.authorAtkins, John F.
dc.contributor.funderEuropean Research Council
dc.contributor.funderWellcome Trust
dc.contributor.funderNational Institutes of Health
dc.contributor.funderScience Foundation Ireland
dc.date.accessioned2017-09-26T11:39:19Z
dc.date.available2017-09-26T11:39:19Z
dc.date.issued2017
dc.description.abstractPositioning test sequences between fused reporters permits monitoring of both translation levels and framing, before and after the test sequence. Many studies, including those on recoding such as productive ribosomal frameshifting and stop codon readthrough, use distinguishable luciferases or fluorescent proteins as reporters. Occasional distortions, due to test sequence product interference with the individual reporter activities or stabilities, are here shown to be avoidable by the introduction of tandem StopGo sequences (2A) flanking the test sequence. Using this new vector system (pSGDluc), we provide evidence for the use of a 3' stem-loop stimulator for ACP2 readthrough, but failed to detect the reported VEGFA readthrough.en
dc.description.sponsorshipNational Institutes of Health (R01GM114291; R21ES022716); Wellcome Trust (106207)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationLoughran, G., Howard, M. T., Firth, A. E. and Atkins, J. F. (2017) 'Avoidance of reporter assay distortions from fused dual reporters', RNA, 23(8) (6pp). doi: 10.1261/rna.061051.117en
dc.identifier.doi10.1261/rna.061051.117
dc.identifier.endpage1289
dc.identifier.issn1355-8382
dc.identifier.issn1469-9001
dc.identifier.issued8
dc.identifier.journaltitleRNAen
dc.identifier.startpage1285
dc.identifier.urihttps://hdl.handle.net/10468/4784
dc.identifier.volume23
dc.language.isoenen
dc.publisherCold Spring Harbor Lab Pressen
dc.relation.projectinfo:eu-repo/grantAgreement/SFI/SFI Investigator Programme/12/IP/1492/IE/Using ribosome profiling to study translation initiation/elongation and facilitate optimization of protein synthesis/
dc.relation.projectinfo:eu-repo/grantAgreement/SFI/SFI Investigator Programme/13/IA/1853/IE/Dynamic redefinition of codons: From antivirals to an essential micronutrient/
dc.relation.projectinfo:eu-repo/grantAgreement/EC/H2020::ERC::ERC-COG/646891/EU/Systematic discovery of functional elements in RNA virus genomes: an Encyclopedia of RNA Virus Elements/ERVE
dc.relation.urihttp://rnajournal.cshlp.org/content/23/8/1285
dc.rights© 2017, Loughran et al. Published by Cold Spring Harbor Laboratory Press for the RNA Society. This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectRecodingen
dc.subjectFrameshiftingen
dc.subjectReadthroughen
dc.subjectDual luciferaseen
dc.subjectStopGoen
dc.subjectVEGF-Axen
dc.subjectCleavageen
dc.subjectStopen
dc.subject2Aen
dc.subjectCodonen
dc.subjectTranslationen
dc.subjectPolyproteinen
dc.subjectAphthovirusen
dc.subjectReadthroughen
dc.subjectFusionen
dc.titleAvoidance of reporter assay distortions from fused dual reportersen
dc.typeArticle (peer-reviewed)en
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