A CRISPR-dCas13 RNA-editing tool to study alternative splicing.

Autor:	Núñez-Álvarez Y; Institut de Génétique Humaine, Université de Montpellier, CNRS UMR9002, Montpellier, France., Espie-Caullet T; Institut de Génétique Humaine, Université de Montpellier, CNRS UMR9002, Montpellier, France.; Institut Curie, Paris-Saclay Research University, CNRS UMR3348, 91401 Orsay, France.; Team supported by la Ligue contre le Cancer, France., Buhagiar G; Institut Curie, Paris-Saclay Research University, CNRS UMR3348, 91401 Orsay, France.; Team supported by la Ligue contre le Cancer, France., Rubio-Zulaika A; Department of Neurosciences, Biogipuzkoa Health Research Institute, 20014 San Sebastián, Spain., Alonso-Marañón J; Department of Neurosciences, Biogipuzkoa Health Research Institute, 20014 San Sebastián, Spain., Luna-Pérez E; Institut Curie, Paris-Saclay Research University, CNRS UMR3348, 91401 Orsay, France.; Team supported by la Ligue contre le Cancer, France., Blazquez L; Department of Neurosciences, Biogipuzkoa Health Research Institute, 20014 San Sebastián, Spain.; Ikerbasque, Basque Foundation for Science, 48009 Bilbao, Spain.; CIBERNED, ISCIII (CIBER, Carlos III Institute, Spanish Ministry of Sciences and Innovation), 28031 Madrid, Spain., Luco RF; Institut de Génétique Humaine, Université de Montpellier, CNRS UMR9002, Montpellier, France.; Institut Curie, Paris-Saclay Research University, CNRS UMR3348, 91401 Orsay, France.; Team supported by la Ligue contre le Cancer, France.
Jazyk:	angličtina
Zdroj:	Nucleic acids research [Nucleic Acids Res] 2024 Oct 28; Vol. 52 (19), pp. 11926-11939.
DOI:	10.1093/nar/gkae682
Abstrakt:	Alternative splicing allows multiple transcripts to be generated from the same gene to diversify the protein repertoire and gain new functions despite a limited coding genome. It can impact a wide spectrum of biological processes, including disease. However, its significance has long been underestimated due to limitations in dissecting the precise role of each splicing isoform in a physiological context. Furthermore, identifying key regulatory elements to correct deleterious splicing isoforms has proven equally challenging, increasing the difficulty of tackling the role of alternative splicing in cell biology. In this work, we take advantage of dCasRx, a catalytically inactive RNA targeting CRISPR-dCas13 ortholog, to efficiently switch alternative splicing patterns of endogenous transcripts without affecting overall gene expression levels cost-effectively. Additionally, we demonstrate a new application for the dCasRx splice-editing system to identify key regulatory RNA elements of specific splicing events. With this approach, we are expanding the RNA toolkit to better understand the regulatory mechanisms underlying alternative splicing and its physiological impact in various biological processes, including pathological conditions. (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)
Databáze:	MEDLINE
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