Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation
- Resource Type
- Authors
- Juana Fernández-Rodríguez; Bernat Gel; Conxi Lázaro; Gabrijela Dumbovic; Meritxell Carrió; Josep Biayna; Helena Mazuelas; Ernest Terribas; Ignacio Blanco; Eduard Serra; Elisabeth Castellanos; Inma Rosas
- Source
- Scientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
Scientific Reports
Dipòsit Digital de la UB
Universidad de Barcelona
r-IGTP. Repositorio Institucional de Producción Científica del Instituto de Investigación Germans Trias i Pujol
instname
- Subject
- MAPK/ERK pathway
Gene isoform
congenital, hereditary, and neonatal diseases and abnormalities
Neurofibromatosis 1
GTPase-activating protein
RNA splicing
Science
Oligonucleotides
PC12 Cells
Article
Neurofibromatosis
Exon
Gene expression
Animals
Humans
Gene
Neurons
Neurofibromatosi
Multidisciplinary
Neurofibromin 1
biology
Chemistry
Alternative splicing
GTPase-Activating Proteins
Oligonucleòtids
Cell Differentiation
Exons
Oligonucleotides, Antisense
Cell biology
Rats
nervous system diseases
Alternative Splicing
Disease Models, Animal
biology.protein
ras Proteins
Medicine
Signal Transduction
Cell signalling
- Language
- English
- ISSN
- 2045-2322
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.