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Biophys Chem. 2009 Aug;143(3):166-9. doi: 10.1016/j.bpc.2009.05.001. Epub 2009 May 12.

Studies of a viral suppressor of RNA silencing p19-CFP fusion protein: a FRET-based probe for sensing double-stranded fluorophore tagged small RNAs.

Author information

1
Steacie Institute for Molecular Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Canada. Roger.Koukiekolo@nrc.ca

Erratum in

  • Biophys Chem. 2009 Nov;145(1):45.

Abstract

Eukaryotes have evolved complex cellular responses to double-stranded RNA. One response that is highly conserved across many species is the RNA silencing pathway. Tombusviruses have evolved a mechanism to evade the RNA silencing pathway that involves a small protein, p19, that acts as a suppressor of RNA silencing. This protein binds specifically to small-interfering RNAs (siRNAs) with nanomolar affinity in a sequence-independent manner and with size selectivity. Here we demonstrate a new approach for rapidly determining the quantities of siRNA using fluorescence resonance energy transfer (FRET) between the Carnation Italian ringspot virus (CIRV) p19-CFP fusion protein and Cy3-labeled siRNA. The CIRV p19 fusion protein binds double-stranded siRNAs with nanomolar affinity as determined by FRET. [corrected].

PMID:
19491057
DOI:
10.1016/j.bpc.2009.05.001
[Indexed for MEDLINE]

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