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RNA. 2014 Jul;20(7):985-8. doi: 10.1261/rna.045054.114. Epub 2014 May 12.

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

Author information

1
Department of Cell Biology Department of Molecular Biophysics and Biochemistry.
2
Department of Molecular Biophysics and Biochemistry Howard 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.

KEYWORDS:

Argonaute; biotin; microRNA

PMID:
24821854
PMCID:
PMC4114695
DOI:
10.1261/rna.045054.114
[Indexed for MEDLINE]
Free PMC Article

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