Paired guide RNA CRISPR-Cas9 screening for protein-coding genes and lncRNAs involved in transdifferentiation of human B-cells to macrophages.
Détails
Télécharger: 35619054_BIB_64A29905BC65.pdf (5448.40 [Ko])
Etat: Public
Version: Final published version
Licence: CC BY 4.0
Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_64A29905BC65
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Paired guide RNA CRISPR-Cas9 screening for protein-coding genes and lncRNAs involved in transdifferentiation of human B-cells to macrophages.
Périodique
BMC genomics
ISSN
1471-2164 (Electronic)
ISSN-L
1471-2164
Statut éditorial
Publié
Date de publication
26/05/2022
Peer-reviewed
Oui
Volume
23
Numéro
1
Pages
402
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: epublish
Publication Status: epublish
Résumé
CRISPR-Cas9 screening libraries have arisen as a powerful tool to identify protein-coding (pc) and non-coding genes playing a role along different processes. In particular, the usage of a nuclease active Cas9 coupled to a single gRNA has proven to efficiently impair the expression of pc-genes by generating deleterious frameshifts. Here, we first demonstrate that targeting the same gene simultaneously with two guide RNAs (paired guide RNAs, pgRNAs) synergistically enhances the capacity of the CRISPR-Cas9 system to knock out pc-genes. We next design a library to target, in parallel, pc-genes and lncRNAs known to change expression during the transdifferentiation from pre-B cells to macrophages. We show that this system is able to identify known players in this process, and also predicts 26 potential novel ones, of which we select four (two pc-genes and two lncRNAs) for deeper characterization. Our results suggest that in the case of the candidate lncRNAs, their impact in transdifferentiation may be actually mediated by enhancer regions at the targeted loci, rather than by the lncRNA transcripts themselves. The CRISPR-Cas9 coupled to a pgRNAs system is, therefore, a suitable tool to simultaneously target pc-genes and lncRNAs for genomic perturbation assays.
Mots-clé
CRISPR-Cas Systems, Cell Transdifferentiation, Humans, Macrophages, RNA, Guide/genetics, RNA, Long Noncoding/genetics, CRISPR-Cas9 screening, Cellular transdifferentiation, Genome editing, Human B-cells, Long non-coding RNA (lncRNA), Paired guide RNA (pgRNA)
Pubmed
Web of science
Open Access
Oui
Création de la notice
17/06/2022 13:15
Dernière modification de la notice
23/01/2024 7:26