ArticleCommunications biology2021
A multiplex CRISPR interference tool for virulence gene interrogation in Legionella pneumophila.
Article in Communications biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Mobile-CRISPRi as a tool for genetic manipulation in the intracellular pathogenApplied and environmental microbiology · 2026Article
- Dual CRISPRi-seq for genome-wide genetic interaction studies identifies key genes involved in the pneumococcal cell cycle.Cell systems · 2025Article
- A programmable, selection-free CRISPR interference system iniScience · 2025Article
- Application of the SpCas9 inhibitor BRD0539 for CRISPR/Cas9-based genetic tools inBioscience of microbiota, food and health · 2025Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- CRISPRi functional genomics in bacteria and its application to medical and industrial research.Microbiology and molecular biology reviews : MMBR · 2024Review
- Legionella pneumophila exploits the endo-lysosomal network for phagosome biogenesis by co-opting SUMOylated Rab7.PLoS pathogens · 2024Article
- Targeted repression ofmBio · 2024Article
- Investigating Pseudomonas aeruginosa Gene Function During Pathogenesis Using Mobile-CRISPRi.Methods in molecular biology (Clifton, N.J.) · 2024Article
- A randomized multiplex CRISPRi-Seq approach for the identification of critical combinations of genes.eLife · 2023Article
- A randomized multiplex CRISPRi-Seq approach for the identification of critical combinations of genes.bioRxiv : the preprint server for biology · 2023Article
- Article
- CRISPRi-mediated tunable control of gene expression level with engineered single-guide RNA in Escherichia coli.Nucleic acids research · 2023Article
- Article
- VpdC is a ubiquitin-activated phospholipase effector that regulatesProceedings of the National Academy of Sciences of the United States of America · 2022Article
- CRISPR-Based Approaches for Gene Regulation in Non-Model Bacteria.Frontiers in genome editing · 2022Review
- Article
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Authors and funding
4 authors.
Funding
Abstract
Catalytically inactive dCas9 imposes transcriptional gene repression by sterically precluding RNA polymerase activity at a given gene to which it was directed by CRISPR (cr)RNAs. This gene silencing technology, known as CRISPR interference (CRISPRi), has been employed in various bacterial species to interrogate genes, mostly individually or in pairs. Here, we developed a multiplex CRISPRi platform in the pathogen Legionella pneumophila capable of silencing up to ten genes simultaneously. Constraints on precursor-crRNA expression were overcome by combining a strong promoter with a boxA element upstream of a CRISPR array. Using crRNAs directed against virulence protein-encoding genes, we demonstrated that CRISPRi is fully functional not only during growth in axenic media, but also during macrophage infection, and that gene depletion by CRISPRi recapitulated the growth defect of deletion strains. By altering the position of crRNA-encoding spacers within the CRISPR array, our platform achieved the gradual depletion of targets that was mirrored by the severity in phenotypes. Multiplex CRISPRi thus holds great promise for probing large sets of genes in bulk in order to decipher virulence strategies of L. pneumophila and other bacterial pathogens.
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