Argonaute HITS-CLIP decodes microRNA-mRNA interaction maps

被引：1330

作者：

Chi, Sung Wook ^{[1
,2
]}

Zang, Julie B. ^{[1
,2
]}

Mele, Aldo ^{[1
,2
]}

Darnell, Robert B. ^{[1
,2
]}

机构：

[1] Rockefeller Univ, Mol Neurooncol Lab, New York, NY 10021 USA

[2] Rockefeller Univ, Howard Hughes Med Inst, New York, NY 10021 USA

来源：

NATURE | 2009年 / 460卷 / 7254期

关键词：

SYSTEMATIC IDENTIFICATION; NEURONAL DIFFERENTIATION; TARGET PREDICTIONS; BINDING PROTEINS; CROSS-LINKING; BRAIN; ELEGANS; CELLS; IMMUNOPRECIPITATION; TRANSLATION;

D O I：

10.1038/nature08170

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

MicroRNAs (miRNAs) have critical roles in the regulation of gene expression; however, as miRNA activity requires base pairing with only 628 nucleotides of messenger RNA, predicting target mRNAs is a major challenge. Recently, high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation (HITS-CLIP) has identified functional protein-RNA interaction sites. Here we use HITS-CLIP to covalently crosslink native argonaute (Ago, also called Eif2c) protein-RNA complexes in mouse brain. This produced two simultaneous data sets-Ago-miRNA and Ago-mRNA binding sites-that were combined with bioinformatic analysis to identify interaction sites between miRNA and target mRNA. We validated genome-wide interaction maps for miR-124, and generated additional maps for the 20 most abundant miRNAs present in P13 mouse brain. Ago HITS-CLIP provides a general platform for exploring the specificity and range of miRNA action in vivo, and identifies precise sequences for targeting clinically relevant miRNA-mRNA interactions.

引用

页码：479 / 486

页数：8

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