Avoidance of reporter assay distortions from fused dual reporters
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Published Version
Date
2017
Authors
Loughran, Gary
Howard, Michael T.
Firth, Andrew E.
Atkins, John F.
Journal Title
Journal ISSN
Volume Title
Publisher
Cold Spring Harbor Lab Press
Published Version
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.
Description
Keywords
Recoding , Frameshifting , Readthrough , Dual luciferase , StopGo , VEGF-Ax , Cleavage , Stop , 2A , Codon , Translation , Polyprotein , Aphthovirus , Readthrough , Fusion
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