Genetic engineering of a temperate phage-based delivery system for CRISPR/Cas9 antimicrobials against Staphylococcus aureus

Discovery of clustered, regularly interspaced, short palindromic repeats and the Cas9 RNA-guided nuclease (CRISPR/Cas9) system provides a new opportunity to create programmable gene-specific antimicrobials that are far less likely to drive resistance than conventional antibiotics. However, the pract...

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Bibliographic Details
Published in:Scientific reports 2017-03, Vol.7 (1), p.44929-44929, Article 44929
Main Authors: Park, Joo Youn, Moon, Bo Youn, Park, Juw Won, Thornton, Justin A., Park, Yong Ho, Seo, Keun Seok
Format: Article
Language:English
Subjects:
42/41
42/70
631/326/1321
631/326/22/1290
631/326/432
Antibiotics
Antimicrobial agents
CRISPR
Genetic engineering
Genomes
Host range
Host specificity
Humanities and Social Sciences
multidisciplinary
Nuclease
Phages
Ribonucleic acid
RNA
Science
Tail fiber protein
Virulence
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Summary:Discovery of clustered, regularly interspaced, short palindromic repeats and the Cas9 RNA-guided nuclease (CRISPR/Cas9) system provides a new opportunity to create programmable gene-specific antimicrobials that are far less likely to drive resistance than conventional antibiotics. However, the practical therapeutic use of CRISPR/Cas9 is still questionable due to current shortcomings in phage-based delivery systems such as inefficient delivery, narrow host range, and potential transfer of virulence genes by generalized transduction. In this study, we demonstrate genetic engineering strategies to overcome these shortcomings by integrating CRISPR/Cas9 system into a temperate phage genome, removing major virulence genes from the host chromosome, and expanding host specificity of the phage by complementing tail fiber protein. This significantly improved the efficacy and safety of CRISPR/Cas9 antimicrobials to therapeutic levels in both in vitro and in vivo assays. The genetic engineering tools and resources established in this study are expected to provide an efficacious and safe CRISPR/Cas9 antimicrobial, broadly applicable to Staphylococcus aureus .
ISSN:2045-2322
2045-2322
DOI:10.1038/srep44929