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Updated: Apr 29, 2026

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
Published on: June 12, 2018
3'-Biotin-tagged microRNA-27 does not associate with Argonaute proteins in cells
Yang Eric Guo1, Joan A Steitz2
1Department of Cell Biology Department of Molecular Biophysics and Biochemistry.
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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