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
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
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