title: 3'-Biotin-tagged microRNA-27 does not associate with Argonaute proteins in cells.
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.
authors.affiliation: Department of Cell Biology Department of Molecular Biophysics and Biochemistry.
authors.forename: Yang Eric
authors.initials: YE
authors.lastname: Guo
authors.affiliation: 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.
authors.forename: Joan A
authors.initials: JA
authors.lastname: Steitz
mesh_terms: D020413:3' Untranslated Regions; D060565:Argonaute Proteins / Q000378:metabolism*; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D035683:MicroRNAs / Q000737:chemistry / Q000378:metabolism*; D015336:Molecular Probe Techniques / Q000592:standards; D011485:Protein Binding; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D060565:Argonaute Proteins; C531388:MIRN27 microRNA, human; D035683:MicroRNAs; D001710:Biotin
keywords: Argonaute; biotin; microRNA