Avoidance of reporter assay distortions from fused dual reporters
dc.contributor.author | Loughran, Gary | |
dc.contributor.author | Howard, Michael T. | |
dc.contributor.author | Firth, Andrew E. | |
dc.contributor.author | Atkins, John F. | |
dc.contributor.funder | European Research Council | |
dc.contributor.funder | Wellcome Trust | |
dc.contributor.funder | National Institutes of Health | |
dc.contributor.funder | Science Foundation Ireland | |
dc.date.accessioned | 2017-09-26T11:39:19Z | |
dc.date.available | 2017-09-26T11:39:19Z | |
dc.date.issued | 2017 | |
dc.description.abstract | Positioning 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.sponsorship | National Institutes of Health (R01GM114291; R21ES022716); Wellcome Trust (106207) | en |
dc.description.status | Peer reviewed | en |
dc.description.version | Published Version | en |
dc.format.mimetype | application/pdf | en |
dc.identifier.citation | Loughran, 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.061051.117 | en |
dc.identifier.doi | 10.1261/rna.061051.117 | |
dc.identifier.endpage | 1289 | |
dc.identifier.issn | 1355-8382 | |
dc.identifier.issn | 1469-9001 | |
dc.identifier.issued | 8 | |
dc.identifier.journaltitle | RNA | en |
dc.identifier.startpage | 1285 | |
dc.identifier.uri | https://hdl.handle.net/10468/4784 | |
dc.identifier.volume | 23 | |
dc.language.iso | en | en |
dc.publisher | Cold Spring Harbor Lab Press | en |
dc.relation.project | info: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.project | info:eu-repo/grantAgreement/SFI/SFI Investigator Programme/13/IA/1853/IE/Dynamic redefinition of codons: From antivirals to an essential micronutrient/ | |
dc.relation.project | info: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.uri | http://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.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.subject | Recoding | en |
dc.subject | Frameshifting | en |
dc.subject | Readthrough | en |
dc.subject | Dual luciferase | en |
dc.subject | StopGo | en |
dc.subject | VEGF-Ax | en |
dc.subject | Cleavage | en |
dc.subject | Stop | en |
dc.subject | 2A | en |
dc.subject | Codon | en |
dc.subject | Translation | en |
dc.subject | Polyprotein | en |
dc.subject | Aphthovirus | en |
dc.subject | Readthrough | en |
dc.subject | Fusion | en |
dc.title | Avoidance of reporter assay distortions from fused dual reporters | en |
dc.type | Article (peer-reviewed) | en |
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