Purification of noncoding RNA and bound proteins using FLAG peptide-conjugated antisense-oligonucleotides

Shungo Adachi; Tohru Natsume

doi:10.1007/978-1-4939-2253-6_16

Purification of noncoding RNA and bound proteins using FLAG peptide-conjugated antisense-oligonucleotides

Methods Mol Biol. 2015:1262:265-74. doi: 10.1007/978-1-4939-2253-6_16.

Authors

Shungo Adachi¹, Tohru Natsume

Affiliation

¹ Molecular Profiling Research Center for Drug Discovery (molprof), National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan.

PMID: 25555587
DOI: 10.1007/978-1-4939-2253-6_16

Abstract

To understand the function of certain RNAs, including noncoding RNAs, it is important to identify the proteins that interact with the RNAs. Here we describe the method for purification of ribonucleoprotein (RNP) complexes composed of specific cellular RNAs by pull-down with FLAG peptide-conjugated antisense oligonucleotide (ASO). Using this method, we identified a novel protein component of U7 snRNP complex.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

HeLa Cells
Humans
Mass Spectrometry / methods*
Oligonucleotides, Antisense / chemistry
Oligopeptides / chemistry
Protein Binding
RNA, Untranslated / isolation & purification*
RNA, Untranslated / metabolism*
Ribonucleoprotein, U7 Small Nuclear / metabolism*

Substances

Oligonucleotides, Antisense
Oligopeptides
RNA, Untranslated
Ribonucleoprotein, U7 Small Nuclear
FLAG peptide