title: Tandem affinity purification of miRNA target mRNAs (TAP-Tar).
abstract: MicroRNAs (miRNAs) bind to Argonaute proteins, and together they form the RISC complex and regulate target mRNA translation and/or stability. Identification of mRNA targets is key to deciphering the physiological functions and mode of action of miRNAs. In mammals, miRNAs are generally poorly homologous to their target sequence, and target identification cannot be based solely on bioinformatics. Here, we describe a biochemical approach, based on tandem affinity purification, in which mRNA/miRNA complexes are sequentially pulled down, first via the Argonaute moiety and then via the miRNA. Our 'TAP-Tar' procedure allows the specific pull down of mRNA targets of miRNA. It is useful for validation of targets predicted in silico, and, potentially, for discovery of previously uncharacterized targets.
authors: CNRS FRE 2944, Institut André Lwoff, Villejuif F-94801 and Université Paris-Sud, Villejuif F-94801, France.
authors: Nora
authors: N
authors: Nonne
authors: Maya
authors: M
authors: Ameyar-Zazoua
authors: Mouloud
authors: M
authors: Souidi
authors: Annick
authors: A
authors: Harel-Bellan
mesh_terms: D019921:Biotinylation; D050687:E2F1 Transcription Factor / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D035683:MicroRNAs / Q000302:isolation & purification* / Q000378:metabolism; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D050687:E2F1 Transcription Factor; C534451:MIRN20a microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger