RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs

RNA–protein interactions frame post-transcriptional regulatory networks and modulate transcription and epigenetics. While the technological advances in RNA sequencing have significantly expanded the repertoire of RNAs, recently developed biochemical approaches combined with sensitive mass-spectromet...

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Main Author: André P. Gerber
Format: Article
Language:English
Published: MDPI AG 2021-02-01
Series:Non-Coding RNA
Subjects:
Online Access:https://www.mdpi.com/2311-553X/7/1/11
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author André P. Gerber
author_facet André P. Gerber
author_sort André P. Gerber
collection DOAJ
description RNA–protein interactions frame post-transcriptional regulatory networks and modulate transcription and epigenetics. While the technological advances in RNA sequencing have significantly expanded the repertoire of RNAs, recently developed biochemical approaches combined with sensitive mass-spectrometry have revealed hundreds of previously unrecognized and potentially novel RNA-binding proteins. Nevertheless, a major challenge remains to understand how the thousands of RNA molecules and their interacting proteins assemble and control the fate of each individual RNA in a cell. Here, I review recent methodological advances to approach this problem through systematic identification of proteins that interact with particular RNAs in living cells. Thereby, a specific focus is given to in vivo approaches that involve crosslinking of RNA–protein interactions through ultraviolet irradiation or treatment of cells with chemicals, followed by capture of the RNA under study with antisense-oligonucleotides and identification of bound proteins with mass-spectrometry. Several recent studies defining interactomes of long non-coding RNAs, viral RNAs, as well as mRNAs are highlighted, and short reference is given to recent in-cell protein labeling techniques. These recent experimental improvements could open the door for broader applications and to study the remodeling of RNA–protein complexes upon different environmental cues and in disease.
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spelling doaj.art-bf60cef9759e443e9ffdb134f29c6fba2023-12-11T17:08:02ZengMDPI AGNon-Coding RNA2311-553X2021-02-01711110.3390/ncrna7010011RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAsAndré P. Gerber0Department of Microbial Sciences, School of Biosciences and Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford GU2 7XH, UKRNA–protein interactions frame post-transcriptional regulatory networks and modulate transcription and epigenetics. While the technological advances in RNA sequencing have significantly expanded the repertoire of RNAs, recently developed biochemical approaches combined with sensitive mass-spectrometry have revealed hundreds of previously unrecognized and potentially novel RNA-binding proteins. Nevertheless, a major challenge remains to understand how the thousands of RNA molecules and their interacting proteins assemble and control the fate of each individual RNA in a cell. Here, I review recent methodological advances to approach this problem through systematic identification of proteins that interact with particular RNAs in living cells. Thereby, a specific focus is given to in vivo approaches that involve crosslinking of RNA–protein interactions through ultraviolet irradiation or treatment of cells with chemicals, followed by capture of the RNA under study with antisense-oligonucleotides and identification of bound proteins with mass-spectrometry. Several recent studies defining interactomes of long non-coding RNAs, viral RNAs, as well as mRNAs are highlighted, and short reference is given to recent in-cell protein labeling techniques. These recent experimental improvements could open the door for broader applications and to study the remodeling of RNA–protein complexes upon different environmental cues and in disease.https://www.mdpi.com/2311-553X/7/1/11RNA–protein interactionnon-coding RNARNA aptamerantisense oligonucleotidesaffinity isolationCRISPR/Cas9
spellingShingle André P. Gerber
RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
Non-Coding RNA
RNA–protein interaction
non-coding RNA
RNA aptamer
antisense oligonucleotides
affinity isolation
CRISPR/Cas9
title RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
title_full RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
title_fullStr RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
title_full_unstemmed RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
title_short RNA-Centric Approaches to Profile the RNA–Protein Interaction Landscape on Selected RNAs
title_sort rna centric approaches to profile the rna protein interaction landscape on selected rnas
topic RNA–protein interaction
non-coding RNA
RNA aptamer
antisense oligonucleotides
affinity isolation
CRISPR/Cas9
url https://www.mdpi.com/2311-553X/7/1/11
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