利用CLIP-seq技术在活细胞中发现全转录组范围的RNA结合蛋白印记。

CLIP-seq to discover transcriptome-wide imprinting of RNA binding proteins in living cells.

作者信息

Saulière Jérôme, Le Hir Hervé

机构信息

Ecole Normale Supérieure, Institut de Biologie de l'ENS (IBENS), Inserm U1024, and CNRS UMR 8197, Paris Cedex 05, 75230, France.

出版信息

Methods Mol Biol. 2015;1296:151-60. doi: 10.1007/978-1-4939-2547-6_14.

DOI:10.1007/978-1-4939-2547-6_14

PMID:25791598

Abstract

UV cross-linking and immunoprecipitation coupled to high-throughput sequencing (CLIP-seq) is used to characterize RNA targets of RNA binding proteins (RBP) in a large scale manner. This powerful method allows the stringent purification of direct RNA binding sites of RBPs in living cells. Here, we describe in detail the protocol we employed to identify RNA targets of the human RNA helicase eIF4AIII.

摘要

紫外线交联免疫沉淀结合高通量测序（CLIP-seq）被用于大规模表征RNA结合蛋白（RBP）的RNA靶点。这种强大的方法能够在活细胞中严格纯化RBP的直接RNA结合位点。在此，我们详细描述了我们用于鉴定人类RNA解旋酶eIF4AIII的RNA靶点的实验方案。