title: Identification of RNA-Protein Interactions Through In Vitro RNA Pull-Down Assays.
abstract: Recent advances in next-generation sequencing have revealed that majority of the human genome is transcribed into long and short RNA (ncRNA) transcripts. Many ncRNAs function by interacting with proteins and forming regulatory complexes. RNA-protein interactions are vital in controlling core cellular processes like transcription and translation. Therefore identifying proteins that interact with ncRNAs is central to deciphering ncRNA functions. Here we describe an RNA-protein pull-down assay, which enables the identification of proteins that interact with an RNA under study. As an example we describe pull-down of proteins interacting with ncRNA XIST, which assists in the recruitment of the polycomb-repressive complex-2 (PRC2) and drives X-chromosomal inactivation.
authors.affiliation: Department of Biochemistry, University of Leicester, Leicester, LE1 9HN, UK.
authors.forename: Claire
authors.initials: C
authors.lastname: Barnes
authors.affiliation: School of Biosciences, University of Birmingham, Birmingham, B15 2TT, UK. a.kanhere@bham.ac.uk.
authors.forename: Aditi
authors.initials: A
authors.lastname: Kanhere
mesh_terms: D053595:Embryonic Stem Cells; D015894:Genome, Human; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D063151:Polycomb Repressive Complex 2 / Q000235:genetics / Q000302:isolation & purification*; D062085:RNA, Long Noncoding / Q000235:genetics / Q000302:isolation & purification*; D016601:RNA-Binding Proteins / Q000235:genetics*; D049951:X Chromosome Inactivation / Q000235:genetics
chemical_list: D062085:RNA, Long Noncoding; D016601:RNA-Binding Proteins; C077885:XIST non-coding RNA; D063151:Polycomb Repressive Complex 2
keywords: Biotin; In vitro transcription; Noncoding RNA; Pull-down; RNA–protein interactions; XIST