Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation

Abstract Neurofibromatosis Type 1 (NF1) is a genetic condition affecting approximately 1:3500 persons worldwide. The NF1 gene codes for neurofibromin protein, a GTPase activating protein (GAP) and a negative regulator of RAS. The NF1 gene undergoes alternative splicing of exon 23a (E23a) that codes...

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Autores principales:	Josep Biayna, Helena Mazuelas, Bernat Gel, Ernest Terribas, Gabrijela Dumbovic, Inma Rosas, Juana Fernández-Rodriguez, Ignacio Blanco, Elisabeth Castellanos, Meritxell Carrió, Conxi Lazaro, Eduard Serra
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7e2ef70d624f46fab52c22933b9b591a
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spelling	oai:doaj.org-article:7e2ef70d624f46fab52c22933b9b591a2021-12-02T14:26:48ZUsing antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation10.1038/s41598-021-83152-w2045-2322https://doaj.org/article/7e2ef70d624f46fab52c22933b9b591a2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83152-whttps://doaj.org/toc/2045-2322Abstract Neurofibromatosis Type 1 (NF1) is a genetic condition affecting approximately 1:3500 persons worldwide. The NF1 gene codes for neurofibromin protein, a GTPase activating protein (GAP) and a negative regulator of RAS. The NF1 gene undergoes alternative splicing of exon 23a (E23a) that codes for 21 amino acids placed at the center of the GAP related domain (GRD). E23a-containing type II neurofibromin exhibits a weaker Ras-GAP activity compared to E23a-less type I isoform. Exon E23a has been related with the cognitive impairment present in NF1 individuals. We designed antisense Phosphorodiamidate Morpholino Oligomers (PMOs) to modulate E23a alternative splicing at physiological conditions of gene expression and tested their impact during PC12 cell line neuronal differentiation. Results show that any dynamic modification of the natural ratio between type I and type II isoforms disturbed neuronal differentiation, altering the proper formation of neurites and deregulating both the MAPK/ERK and cAMP/PKA signaling pathways. Our results suggest an opposite regulation of these pathways by neurofibromin and the possible existence of a feedback loop sensing neurofibromin-related signaling. The present work illustrates the utility of PMOs to study alternative splicing that could be applied to other alternatively spliced genes in vitro and in vivo.Josep BiaynaHelena MazuelasBernat GelErnest TerribasGabrijela DumbovicInma RosasJuana Fernández-RodriguezIgnacio BlancoElisabeth CastellanosMeritxell CarrióConxi LazaroEduard SerraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Josep Biayna Helena Mazuelas Bernat Gel Ernest Terribas Gabrijela Dumbovic Inma Rosas Juana Fernández-Rodriguez Ignacio Blanco Elisabeth Castellanos Meritxell Carrió Conxi Lazaro Eduard Serra Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
description	Abstract Neurofibromatosis Type 1 (NF1) is a genetic condition affecting approximately 1:3500 persons worldwide. The NF1 gene codes for neurofibromin protein, a GTPase activating protein (GAP) and a negative regulator of RAS. The NF1 gene undergoes alternative splicing of exon 23a (E23a) that codes for 21 amino acids placed at the center of the GAP related domain (GRD). E23a-containing type II neurofibromin exhibits a weaker Ras-GAP activity compared to E23a-less type I isoform. Exon E23a has been related with the cognitive impairment present in NF1 individuals. We designed antisense Phosphorodiamidate Morpholino Oligomers (PMOs) to modulate E23a alternative splicing at physiological conditions of gene expression and tested their impact during PC12 cell line neuronal differentiation. Results show that any dynamic modification of the natural ratio between type I and type II isoforms disturbed neuronal differentiation, altering the proper formation of neurites and deregulating both the MAPK/ERK and cAMP/PKA signaling pathways. Our results suggest an opposite regulation of these pathways by neurofibromin and the possible existence of a feedback loop sensing neurofibromin-related signaling. The present work illustrates the utility of PMOs to study alternative splicing that could be applied to other alternatively spliced genes in vitro and in vivo.
format	article
author	Josep Biayna Helena Mazuelas Bernat Gel Ernest Terribas Gabrijela Dumbovic Inma Rosas Juana Fernández-Rodriguez Ignacio Blanco Elisabeth Castellanos Meritxell Carrió Conxi Lazaro Eduard Serra
author_facet	Josep Biayna Helena Mazuelas Bernat Gel Ernest Terribas Gabrijela Dumbovic Inma Rosas Juana Fernández-Rodriguez Ignacio Blanco Elisabeth Castellanos Meritxell Carrió Conxi Lazaro Eduard Serra
author_sort	Josep Biayna
title	Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
title_short	Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
title_full	Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
title_fullStr	Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
title_full_unstemmed	Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
title_sort	using antisense oligonucleotides for the physiological modulation of the alternative splicing of nf1 exon 23a during pc12 neuronal differentiation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/7e2ef70d624f46fab52c22933b9b591a
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Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation

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