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Analysis of Protein–RNA Interactions with Single‐Nucleotide Resolution Using iCLIP and Next‐Generation Sequencing

2011/12/14 by Julian König, Nicholas J. McGlincy, Jernej Ule · 1 citation
Biochemistry, Genetics and Molecular Biology · #RNA and protein synthesis mechanisms #RNA Research and Splicing #RNA modifications and cancer #Computational biology #RNA-binding protein #RNA #Nucleotide #Biology #Gene #Binding site #Genetics

paper · doi:10.1002/9783527644582.ch10

openalex publication_date 2011/12/14 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/19

Abstract

Post-transcriptional regulation of gene expression is controlled by the unique composition and spatial arrangement of RNA-binding proteins (RBPs) on individual transcripts. Therefore, understanding post-transcriptional regulation requires precise and comprehensive binding site maps for RBPs. UV cross-linking and immunoprecipitation (CLIP) is a state-of-the-art technique for generating such maps on a genome-wide scale. However, data complexity is often limited and the resolution of the resulting maps is confined to approximately 30 nucleotides. This, in turn, complicates the identification of individual binding sites. We recently described individual-nucleotide resolution CLIP (iCLIP) – an approach that both increases data complexity and allows binding site detection at single-nucleotide resolution. Here, we present the latest version of our iCLIP protocol, discussing critical aspects and recent modifications.

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