Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation.
Alternative Splicing
/ drug effects
Animals
Cell Differentiation
/ drug effects
Disease Models, Animal
Exons
/ genetics
GTPase-Activating Proteins
/ genetics
Humans
Neurofibromatosis 1
/ genetics
Neurofibromin 1
/ genetics
Neurons
/ cytology
Oligonucleotides, Antisense
/ genetics
PC12 Cells
Rats
Signal Transduction
/ drug effects
ras Proteins
/ genetics
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
11 02 2021
11 02 2021
Historique:
received:
08
05
2020
accepted:
29
01
2021
entrez:
12
2
2021
pubmed:
13
2
2021
medline:
16
11
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 33574490
doi: 10.1038/s41598-021-83152-w
pii: 10.1038/s41598-021-83152-w
pmc: PMC7878752
doi:
Substances chimiques
GTPase-Activating Proteins
0
Neurofibromin 1
0
Oligonucleotides, Antisense
0
ras Proteins
EC 3.6.5.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3661Références
Wiley Interdiscip Rev RNA. 2020 Jul;11(4):e1585
pubmed: 31922356
Cell. 1990 Nov 16;63(4):843-9
pubmed: 2121370
Science. 1997 May 2;276(5313):795-8
pubmed: 9115204
Mol Cell Biol. 2014 Jun;34(12):2188-97
pubmed: 24710274
Eur J Hum Genet. 2004 Feb;12(2):139-49
pubmed: 14560314
Nat Med. 2005 Apr;11(4):440-5
pubmed: 15750600
Biol Chem. 2016 Mar;397(3):215-22
pubmed: 26760308
J Neurocytol. 1998 Dec;27(12):939-52
pubmed: 10659684
PLoS Comput Biol. 2006 Jan;2(1):e4
pubmed: 16424921
Genesis. 2001 Jul;30(3):94-102
pubmed: 11477682
Mol Ther. 2019 Nov 6;27(11):2005-2017
pubmed: 31416775
Sci STKE. 2007 Apr 17;2007(382):pe15
pubmed: 17440132
Clin Sci (Lond). 1994 Nov;87(5):481-5
pubmed: 7874833
Oncogene. 1991 Sep;6(9):1555-9
pubmed: 1923522
Hum Mol Genet. 1995 May;4(5):915-20
pubmed: 7633452
Nat Neurosci. 2002 Feb;5(2):95-6
pubmed: 11788835
Biochem Biophys Res Commun. 1992 Aug 31;187(1):332-9
pubmed: 1520317
Ann N Y Acad Sci. 2005 Nov;1058:62-75
pubmed: 16394126
Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424-8
pubmed: 1065897
BMC Genomics. 2010 Jan 29;11:75
pubmed: 20113474
Hum Mutat. 2012 Jan;33(1):81-5
pubmed: 21922594
Dev Biol. 1994 Feb;161(2):538-51
pubmed: 8314000
Mol Cell Biol. 2008 Feb;28(4):1240-51
pubmed: 18086893
Nucleic Acids Res. 2010 Jan;38(1):253-64
pubmed: 19854948
BMC Mol Biol. 2012 Dec 10;13:35
pubmed: 23227900
J Vis Exp. 2018 Aug 26;(138):
pubmed: 30199013
J Biol Chem. 2003 Jul 18;278(29):26958-69
pubmed: 12730209
Biosci Rep. 2012 Apr 1;32(2):131-8
pubmed: 22115364
Gene Ther. 2006 Oct;13(19):1373-81
pubmed: 16724091
Dev Biol. 2014 Sep 1;393(1):24-32
pubmed: 25014653
Methods Mol Biol. 2017;1648:39-52
pubmed: 28766288
Mol Cell Biol. 1993 Jan;13(1):487-95
pubmed: 8417346
Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8673-7
pubmed: 8378346
Nucleic Acid Ther. 2017 Jun;27(3):130-143
pubmed: 28118087
Neurotoxicol Teratol. 2004 May-Jun;26(3):397-406
pubmed: 15113601
Exp Cell Res. 1996 Oct 10;228(1):44-9
pubmed: 8892969
Hum Mol Genet. 2017 Oct 1;26(19):3797-3807
pubmed: 28934393
Cell. 2008 Oct 31;135(3):549-60
pubmed: 18984165
Hum Mutat. 2011 Jul;32(7):705-9
pubmed: 21394830
Hum Gene Ther. 2013 Mar;24(3):331-42
pubmed: 23339722
Science. 2002 May 31;296(5573):1648-9
pubmed: 12040181
Cell. 1995 Jan 27;80(2):179-85
pubmed: 7834738
Nat Genet. 2001 Apr;27(4):399-405
pubmed: 11279521
Biochem Biophys Res Commun. 1991 Dec 31;181(3):955-61
pubmed: 1662505
Nat Rev Cancer. 2015 May;15(5):290-301
pubmed: 25877329
Cell. 1990 Aug 10;62(3):599-608
pubmed: 2116237
J Virol. 2011 Mar;85(5):2463-8
pubmed: 21159861
Nature. 2000 Feb 24;403(6772):895-8
pubmed: 10706287
Cell. 1990 Nov 16;63(4):851-9
pubmed: 2121371
Nucleic Acids Res. 2001 Oct 1;29(19):3965-74
pubmed: 11574678
Cytometry A. 2004 Apr;58(2):167-76
pubmed: 15057970
Hum Mutat. 2009 Mar;30(3):454-62
pubmed: 19241459
Genome Res. 2012 Jan;22(1):35-50
pubmed: 21974994
Nature. 2002 Jan 31;415(6871):526-30
pubmed: 11793011
PLoS One. 2008 Jul 30;3(7):e2806
pubmed: 18665247
Trends Cell Biol. 2002 Jun;12(6):258-66
pubmed: 12074885
Hum Mol Genet. 2000 Dec 12;9(20):3055-64
pubmed: 11115850
FEBS J. 2005 Jul;272(14):3491-504
pubmed: 16008550
Biomed Pharmacother. 1994;48(8-9):365-72
pubmed: 7858173
J Neurosci Res. 1993 Oct 1;36(2):216-23
pubmed: 7505343
Nature. 1991 Jun 13;351(6327):576-9
pubmed: 1904555
Nat Genet. 1997 Mar;15(3):281-4
pubmed: 9054942
Arch Dermatol Res. 1995;287(5):413-6
pubmed: 7625848
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
J Med Genet. 2007 Feb;44(2):81-8
pubmed: 17105749
J Invest Dermatol. 2009 Oct;129(10):2497-506
pubmed: 19369967
Cell Growth Differ. 1995 Mar;6(3):315-23
pubmed: 7794799
Hum Mol Genet. 2004 Oct 15;13(20):2409-20
pubmed: 15333583
Mol Carcinog. 1992;6(2):83-7
pubmed: 1388685
Eur J Hum Genet. 2013 Jul;21(7):769-73
pubmed: 23188051
PLoS One. 2013 Apr 22;8(4):e61472
pubmed: 23630592
Trends Genet. 2013 Aug;29(8):442-8
pubmed: 23648015