Development of sequence-specific antimicrobials based on programmable CRISPR-Cas nucleases

By David Bikard, Chad W. Euler, Wenyan Jiang, P. Nussenzweig, Gregory W. Goldberg, X. Duportet, V. Fischetti, L. Marraffini. Nature Biotechnology, 2014

799 citations recorded by the source corpus.

Antibiotics target conserved bacterial cellular pathways or growth functions and therefore cannot selectively kill specific members of a complex microbial population. Here, we develop programmable, sequence-specific antimicrobials using the RNA-guided nuclease Cas9 (refs.1,2) delivered by a bacteriophage. We show that Cas9, reprogrammed to target virulence genes, kills virulent, but not avirulent, Staphylococcus aureus. Reprogramming the nuclease to target antibiotic resistance genes destroys staphylococcal plasmids that harbor antibiotic resistance genes and immunizes avirulent staphylococci to prevent the spread of plasmid-borne resistance genes. We also show that CRISPR-Cas9 antimicrobials function in vivo to kill S. aureus in a mouse skin colonization model. This technology creates opportunities to manipulate complex bacterial populations in a sequence-specific manner.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/910132a96da298ba4667f5079454f7e1738bb6ad

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Genetics

1. Ole Lund
2. Rodolphe Barrangou
3. Intawat Nookaew
4. Se-Ran Jun
5. Timothy K. Lu
6. Loren J. Hauser

In Molecular Biology

1. S. Lemire
2. Hiroki Ando
3. Y. Taur
4. Ole Lund
5. Min Wu
6. Rodolphe Barrangou
7. David Wayne Ussery
8. Intawat Nookaew

Citations in the corpus

• The effects of antibiotics on the microbiome throughout development and alternative approaches for therapeutic modulation (2016, background): “Recently, the natural transformation capacity of phages has been coupled with programmable nucleases to enable the generation of phages that specifically kill bacteria with undesirable genomic sequences, such as antibiotic resistance genes or virulence factors [164, 165].”
• Antibiotic-Induced Changes in the Intestinal Microbiota and Disease. (2016, background): “Finally, a strategy to eliminate selected bacterial targets exploiting the CRISPR/Cas9 system was recently proposed [127,128].”
• Insights from 20 years of bacterial genome sequencing (2015, background): “As the role in virulence is elucidated, the Cas9 protein of the system is showing promise as tool for genetically engineering new weapons in the war against antibioticresistant bacteria (Birkard et al. 2014).”
• Pseudomonas aeruginosa: pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics (2022, background): “Creating a synthetic phage with desired characteristics and genomic content would be a creative approach.(355) Recently, mutations in phage host-range-determining regions (HRDRs) to cope with resistance mutation of bacteria have been implemented as an effective method to prevent Escherichia coli inf”
• Reprogramming Bacteriophage Host Range through Structure-Guided Design of Chimeric Receptor Binding Proteins. (2019, background): “For example, several studies have demonstrated that targeted phage vectors can be utilized to re-sensitize or remove antibiotic-resistant bacteria frommixed populations (Bikard et al., 2014; Citorik et al., 2014; Edgar et al., 2012; Goren et al., 2017; Yosef et al., 2015).”
• Identifying and visualizing functional PAM diversity across CRISPR-Cas systems (2016, background): “The programmable and multiplexable nature of DNA/RNA targeting has led to a myriad of applications ranging from gene therapy to antimicrobials (Bikard et al., 2014; Citorik et al., 2014; Cong et al., 2013; Gomaa et al., 2014; Hilton et al., 2015; Mali et al., 2013).”
• Engineering Phage Host-Range and Suppressing Bacterial Resistance through Phage Tail Fiber Mutagenesis (2019, background): “A resurging approach gaining significant interest is phage therapy, in which bacteriophages are used as antimicrobials (Bikard et al., 2014; Chen et al., 2014; Citorik et al., 2014; Devlin et al., 2016; Kutateladze and Adamia, 2010; Kutter et al., 2010, 2015; Lu and Collins, 2007, 2009; Maynard et a”