CRISPRi-TnSeq maps genome-wide interactions between essential and non-essential genes in bacteria.

Détails

ID Serval
serval:BIB_A51A94333B3E
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
CRISPRi-TnSeq maps genome-wide interactions between essential and non-essential genes in bacteria.
Périodique
Nature microbiology
Auteur⸱e⸱s
Jana B., Liu X., Dénéréaz J., Park H., Leshchiner D., Liu B., Gallay C., Zhu J., Veening J.W., van Opijnen T.
ISSN
2058-5276 (Electronic)
ISSN-L
2058-5276
Statut éditorial
Publié
Date de publication
09/2024
Peer-reviewed
Oui
Volume
9
Numéro
9
Pages
2395-2409
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
Genetic interactions identify functional connections between genes and pathways, establishing gene functions or druggable targets. Here we use CRISPRi-TnSeq, CRISPRi-mediated knockdown of essential genes alongside TnSeq-mediated knockout of non-essential genes, to map genome-wide interactions between essential and non-essential genes in Streptococcus pneumoniae. Transposon-mutant libraries constructed in 13 CRISPRi strains enabled screening of ~24,000 gene pairs. This identified 1,334 genetic interactions, including 754 negative and 580 positive interactions. Network analyses show that 17 non-essential genes pleiotropically interact with more than half the essential genes tested. Validation experiments confirmed that a 7-gene subset protects against perturbations. Furthermore, we reveal hidden redundancies that compensate for essential gene loss, relationships between cell wall synthesis, integrity and cell division, and show that CRISPRi-TnSeq identifies synthetic and suppressor-type relationships between both functionally linked and disparate genes and pathways. Importantly, in species where CRISPRi and Tn-Seq are established, CRISPRi-TnSeq should be straightforward to implement.
Mots-clé
Streptococcus pneumoniae/genetics, Streptococcus pneumoniae/metabolism, Genes, Essential/genetics, Genome, Bacterial/genetics, DNA Transposable Elements/genetics, CRISPR-Cas Systems, Gene Regulatory Networks, Clustered Regularly Interspaced Short Palindromic Repeats/genetics, Genes, Bacterial/genetics, Bacterial Proteins/genetics, Bacterial Proteins/metabolism, Gene Knockout Techniques
Pubmed
Web of science
Open Access
Oui
Création de la notice
26/07/2024 12:56
Dernière modification de la notice
10/09/2024 6:17
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