3′-Biotin-tagged microRNA-27 does not associate with Argonaute proteins in cells

Yang Eric Guo; Joan A. Steitz

doi:10.1261/rna.045054.114

3′-Biotin-tagged microRNA-27 does not associate with Argonaute proteins in cells

Yang Eric Guo1,2 and
Joan A. Steitz2,3,4

¹Department of Cell Biology
²Department of Molecular Biophysics and Biochemistry
³Howard Hughes Medical Institute, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06536, USA

Abstract

Synthetic 3′-biotin-tagged microRNAs (miRNAs) have often been used to select interacting messenger RNA (mRNA) and noncoding RNA (ncRNA) targets. Here, we examined the extent of association of 3′-end biotinylated miR-27 with Argonaute (Ago) proteins in transfected human cells using a coimmunoprecipitation assay followed by Northern blot analysis. We report that biotinylated miR-27 does not efficiently associate with Ago compared to unmodified miR-27. These results suggest that 3′-end biotin-modified miRNAs are questionable monitors of miRNA function in cells.

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Footnotes

↵4 Corresponding author

E-mail joan.steitz{at}yale.edu
Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.045054.114.

Received March 2, 2014.
Accepted April 2, 2014.

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