A Novel CRISPR Interference Effector Enabling Functional Gene Characterization with Synthetic Guide RNAs.
Journal
The CRISPR journal
ISSN: 2573-1602
Titre abrégé: CRISPR J
Pays: United States
ID NLM: 101738191
Informations de publication
Date de publication:
Dec 2022
Dec 2022
Historique:
pubmed:
19
10
2022
medline:
16
12
2022
entrez:
18
10
2022
Statut:
ppublish
Résumé
While CRISPR interference (CRISPRi) systems have been widely implemented in pooled lentiviral screening, there has been limited use with synthetic guide RNAs for the complex phenotypic readouts enabled by experiments in arrayed format. Here we describe a novel deactivated Cas9 fusion protein, dCas9-SALL1-SDS3, which produces greater target gene repression than first or second generation CRISPRi systems when used with chemically modified synthetic single guide RNAs (sgRNAs), while exhibiting high target specificity. We show that dCas9-SALL1-SDS3 interacts with key members of the histone deacetylase and Swi-independent three complexes, which are the endogenous functional effectors of SALL1 and SDS3. Synthetic sgRNAs can also be used with
Identifiants
pubmed: 36257604
doi: 10.1089/crispr.2022.0056
pmc: PMC9805873
doi:
Substances chimiques
CRISPR-Associated Protein 9
EC 3.1.-
RNA, Guide, CRISPR-Cas Systems
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
769-786Références
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