CRISPR artificial splicing factors.
Alternative Splicing
Base Sequence
CRISPR-Cas Systems
/ genetics
Cell Line
Cell Line, Tumor
Exons
/ genetics
Fibroblasts
/ metabolism
HEK293 Cells
HeLa Cells
Humans
Muscular Atrophy, Spinal
/ genetics
RNA
/ genetics
RNA Isoforms
/ genetics
RNA Splicing Factors
/ genetics
Survival of Motor Neuron 2 Protein
/ genetics
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
12 06 2020
12 06 2020
Historique:
received:
11
12
2018
accepted:
22
05
2020
entrez:
14
6
2020
pubmed:
14
6
2020
medline:
25
8
2020
Statut:
epublish
Résumé
Alternative splicing allows expression of mRNA isoforms from a single gene, expanding the diversity of the proteome. Its prevalence in normal biological and disease processes warrant precise tools for modulation. Here we report the engineering of CRISPR Artificial Splicing Factors (CASFx) based on RNA-targeting CRISPR-Cas systems. We show that simultaneous exon inclusion and exclusion can be induced at distinct targets by differential positioning of CASFx. We also create inducible CASFx (iCASFx) using the FKBP-FRB chemical-inducible dimerization domain, allowing small molecule control of alternative splicing. Finally, we demonstrate the activation of SMN2 exon 7 splicing in spinal muscular atrophy (SMA) patient fibroblasts, suggesting a potential application of the CASFx system.
Identifiants
pubmed: 32532987
doi: 10.1038/s41467-020-16806-4
pii: 10.1038/s41467-020-16806-4
pmc: PMC7293279
doi:
Substances chimiques
RNA Isoforms
0
RNA Splicing Factors
0
SMN2 protein, human
0
Survival of Motor Neuron 2 Protein
0
RNA
63231-63-0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2973Subventions
Organisme : NCI NIH HHS
ID : P30 CA034196
Pays : United States
Organisme : NHGRI NIH HHS
ID : R01 HG009900
Pays : United States
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