Effective RNA Knockdown Using CRISPR-Cas13a and Molecular Targeting of the
CRISPR-Cas13a
EML4-ALK
RNA knockdown
RNA stability
cell viability
firefly luciferase
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
24 Nov 2020
24 Nov 2020
Historique:
received:
28
09
2020
revised:
20
11
2020
accepted:
20
11
2020
entrez:
1
12
2020
pubmed:
2
12
2020
medline:
6
3
2021
Statut:
epublish
Résumé
RNAi technology has significant potential as a future therapeutic and could theoretically be used to knock down disease-specific RNAs. However, due to frequent off-target effects, low efficiency, and limited accessibility of nuclear transcripts, the clinical application of the technology remains challenging. In this study, we first assessed the stability of Cas13a mRNA and guide RNA. Next, we titrated Cas13a and guide RNA vectors to achieve effective knockdown of firefly luciferase (FLuc) RNA, used as a target transcript. The interference specificity of Cas13a on guide RNA design was next explored. Subsequently, we targeted the
Identifiants
pubmed: 33255340
pii: ijms21238904
doi: 10.3390/ijms21238904
pmc: PMC7727695
pii:
doi:
Substances chimiques
EML4-ALK fusion protein, human
0
Oncogene Proteins, Fusion
0
RNA
63231-63-0
Caspases
EC 3.4.22.-
caspase 13
EC 3.4.22.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Japan Society for the Promotion of Science
ID : 17H02204 and 18K19288
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