2023
DOI: 10.1021/acssynbio.2c00465
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P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri

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 t… Show more

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Cited by 12 publications
(7 citation statements)
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“…The zebrafish larva is a powerful non-mammalian vertebrate model that has been instrumental in the study of the innate immune response to bacterial infection ( Gomes and Mostowy, 2020 ; Stream and Madigan, 2022 ; Torraca and Mostowy, 2018 ). We have previously used S. flexneri infection of zebrafish to study bacterial autophagy ( Mostowy et al, 2013 ), predator-prey interactions ( Willis et al, 2016 ), P1-bacteriophage-mediated immunity ( Huan et al, 2023 ), septin-mediated immunity ( Mazon-Moya et al, 2017 ; Torraca et al, 2023a ; Van Ngo et al, 2023 ), innate immune training ( Gomes et al, 2023 ; Willis et al, 2018 ), Shigella speciation ( Torraca et al, 2019 ) and Shigella persistent infection ( Torraca et al, 2023b ). Here, using RNA-seq, we transcriptionally capture the host response to S. flexneri infection over time.…”
Section: Discussionmentioning
confidence: 99%
“…The zebrafish larva is a powerful non-mammalian vertebrate model that has been instrumental in the study of the innate immune response to bacterial infection ( Gomes and Mostowy, 2020 ; Stream and Madigan, 2022 ; Torraca and Mostowy, 2018 ). We have previously used S. flexneri infection of zebrafish to study bacterial autophagy ( Mostowy et al, 2013 ), predator-prey interactions ( Willis et al, 2016 ), P1-bacteriophage-mediated immunity ( Huan et al, 2023 ), septin-mediated immunity ( Mazon-Moya et al, 2017 ; Torraca et al, 2023a ; Van Ngo et al, 2023 ), innate immune training ( Gomes et al, 2023 ; Willis et al, 2018 ), Shigella speciation ( Torraca et al, 2019 ) and Shigella persistent infection ( Torraca et al, 2023b ). Here, using RNA-seq, we transcriptionally capture the host response to S. flexneri infection over time.…”
Section: Discussionmentioning
confidence: 99%
“…Our work also showed growth advantage of EcN expressing Cas3-Cascade over that expressing Cas9. Although the type II CRISPR-Cas9 system is toxic to many host bacteria, previous work applied this system in killing pathogens or eliminating ARGs 15, 17, 20, 63 . Whereas, an excellent probiotic should not only exert a beneficial effect, but also out-compete other bacteria in the complex ecosystem of the gut microbiome.…”
Section: Discussionmentioning
confidence: 99%
“…Our results indicate that antibiotics are working with the immune system in an additive manner to control S. flexneri infection. These results after the use of antibiotics can be compared to those after the use of the P1 bacteriophage ( Huan et al, 2022 ) or predatory bacteria ( Bdellovibrio bacteriovorus ) to treat against S. flexneri infection in zebrafish ( Willis et al, 2016 ). In the latter case, injection of Bdellovibrio triggers neutrophil recruitment and promotes direct interaction of Bdellovibrio with S. flexneri , whereas antibiotics diluted into the bath water require time to diffuse through the larvae to reach the bacteria.…”
Section: Discussionmentioning
confidence: 99%