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      PureCLIP: capturing target-specific protein–RNA interaction footprints from single-nucleotide CLIP-seq data

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          Abstract

          The iCLIP and eCLIP techniques facilitate the detection of protein–RNA interaction sites at high resolution, based on diagnostic events at crosslink sites. However, previous methods do not explicitly model the specifics of iCLIP and eCLIP truncation patterns and possible biases. We developed PureCLIP ( https://github.com/skrakau/PureCLIP), a hidden Markov model based approach, which simultaneously performs peak-calling and individual crosslink site detection. It explicitly incorporates a non-specific background signal and, for the first time, non-specific sequence biases. On both simulated and real data, PureCLIP is more accurate in calling crosslink sites than other state-of-the-art methods and has a higher agreement across replicates.

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          The online version of this article (doi:10.1186/s13059-017-1364-2) contains supplementary material, which is available to authorized users.

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            Rbfox proteins regulate alternative mRNA splicing through evolutionarily conserved RNA bridges.

            Alternative splicing (AS) enables programmed diversity of gene expression across tissues and development. We show here that binding in distal intronic regions (>500 nucleotides (nt) from any exon) by Rbfox splicing factors important in development is extensive and is an active mode of splicing regulation. Similarly to exon-proximal sites, distal sites contain evolutionarily conserved GCATG sequences and are associated with AS activation and repression upon modulation of Rbfox abundance in human and mouse experimental systems. As a proof of principle, we validated the activity of two specific Rbfox enhancers in KIF21A and ENAH distal introns and showed that a conserved long-range RNA-RNA base-pairing interaction (an RNA bridge) is necessary for Rbfox-mediated exon inclusion in the ENAH gene. Thus we demonstrate a previously unknown RNA-mediated mechanism for AS control by distally bound RNA-binding proteins.
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              HITS-CLIP: panoramic views of protein-RNA regulation in living cells.

              The study of gene regulation in cells has recently begun to shift from a period dominated by the study of transcription factor-DNA interactions to a new focus on RNA regulation. This was sparked by the still-emerging recognition of the central role for RNA in cellular complexity emanating from the RNA World hypothesis, and has been facilitated by technologic advances, in particular high throughput RNA sequencing and crosslinking methods (RNA-Seq, CLIP, and HITS-CLIP). This study will place these advances in context, and, focusing on CLIP, will explain the method, what it can be used for, and how to approach using it. Examples of the successes, limitations, and future of the technique will be discussed. Copyright © 2010 John Wiley & Sons, Ltd.
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                Author and article information

                Contributors
                krakau@molgen.mpg.de
                Journal
                Genome Biol
                Genome Biol
                Genome Biology
                BioMed Central (London )
                1474-7596
                1474-760X
                28 December 2017
                28 December 2017
                2017
                : 18
                : 240
                Affiliations
                [1 ]ISNI 0000 0000 9071 0620, GRID grid.419538.2, Max Planck Institute for Molecular Genetics, ; Ihnestrasse 63–73, Berlin, 14195 Germany
                [2 ]ISNI 0000 0001 2308 1657, GRID grid.462844.8, Sorbonne Universités, ; UPMC Univ Paris 06, CNRS, IBPS, UMR 7238, Laboratoire de Biologie Computationnelle et Quantitative (LCQB), 4 place Jussieu, Paris, 75005 France
                [3 ]ISNI 0000 0000 9116 4836, GRID grid.14095.39, Freie Universität Berlin, ; Takustr. 9, Berlin, 14195 Germany
                Author information
                http://orcid.org/0000-0003-0603-7907
                Article
                1364
                10.1186/s13059-017-1364-2
                5746957
                29284540
                f9a07999-19d0-45be-99d6-2060d5cbeab0
                © The Author(s) 2017

                Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 2 June 2017
                : 24 November 2017
                Categories
                Method
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                © The Author(s) 2017

                Genetics
                protein–rna interaction,iclip-seq,eclip-seq,crosslink sites,hidden markov model
                Genetics
                protein–rna interaction, iclip-seq, eclip-seq, crosslink sites, hidden markov model

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