PUBLICATION
P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri
- Authors
- Huan, Y.W., Torraca, V., Brown, R., Fa-Arun, J., Miles, S.L., Oyarzún, D.A., Mostowy, S., Wang, B.
- ID
- ZDB-PUB-230223-42
- Date
- 2023
- Source
- ACS synthetic biology 12(3): 709-721 (Journal)
- Registered Authors
- Mostowy, Serge, Torraca, Vincenzo
- Keywords
- CRISPR-Cas9, P1 bacteriophage, Shigella flexneri, antimicrobial, phagemid
- MeSH Terms
-
- Animals
- Anti-Infective Agents*
- Bacteriophage P1/genetics
- CRISPR-Cas Systems*/genetics
- Gene Editing
- Shigella flexneri/genetics
- Zebrafish/genetics
- PubMed
- 36802585 Full text @ ACS Synth Biol
Citation
Huan, Y.W., Torraca, V., Brown, R., Fa-Arun, J., Miles, S.L., Oyarzún, D.A., Mostowy, S., Wang, B. (2023) P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri. ACS synthetic biology. 12(3):709-721.
Abstract
The discovery of clustered, regularly interspaced, short palindromic repeats (CRISPR) and the Cas9 RNA-guided nuclease provides unprecedented opportunities to selectively kill specific populations or species of bacteria. However, the use of CRISPR-Cas9 to clear bacterial infections in vivo is hampered by the inefficient delivery of cas9 genetic constructs into bacterial cells. Here, we use a broad-host-range P1-derived phagemid to deliver the CRISPR-Cas9 chromosomal-targeting system into Escherichia coli and the dysentery-causing Shigella flexneri to achieve DNA sequence-specific killing of targeted bacterial cells. We show that genetic modification of the helper P1 phage DNA packaging site (pac) significantly enhances the purity of packaged phagemid and improves the Cas9-mediated killing of S. flexneri cells. We further demonstrate that P1 phage particles can deliver chromosomal-targeting cas9 phagemids into S. flexneriin vivo using a zebrafish larvae infection model, where they significantly reduce the bacterial load and promote host survival. Our study highlights the potential of combining P1 bacteriophage-based delivery with the CRISPR chromosomal-targeting system to achieve DNA sequence-specific cell lethality and efficient clearance of bacterial infection.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping