Avoidance of reporter assay distortions from fused dual reporters

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Avoidance of reporter assay distortions from fused dual reporters

Loughran, Gary; Howard, Michael T.; Firth, Andrew E.; Atkins, John F.
Date: 2017
Copyright: © 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/.
Copyright information: https://creativecommons.org/licenses/by/4.0/
Related: http://rnajournal.cshlp.org/content/23/8/1285
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
Published Version: 10.1261/rna.061051.117

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.

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© 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/. Except where otherwise noted, this item's license is described as © 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/.
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