title: Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials.
abstract: 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.
authors.affiliation: 1] Laboratory of Bacteriology, The Rockefeller University, New York, New York, USA. [2] Present address: Synthetic Biology Group, Institut Pasteur, Paris, France.
authors.forename: David
authors.identifier: 0000000257291211
authors.initials: D
authors.lastname: Bikard
authors.affiliation: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, New York, USA.
authors.forename: Chad W
authors.initials: CW
authors.lastname: Euler
authors.affiliation: Laboratory of Bacteriology, The Rockefeller University, New York, New York, USA.
authors.forename: Wenyan
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Laboratory of Bacteriology, The Rockefeller University, New York, New York, USA.
authors.forename: Philip M
authors.initials: PM
authors.lastname: Nussenzweig
authors.affiliation: Laboratory of Bacteriology, The Rockefeller University, New York, New York, USA.
authors.forename: Gregory W
authors.initials: GW
authors.lastname: Goldberg
authors.affiliation: 1] INRIA Paris-Rocquencourt, Rocquencourt, France. [2] Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Xavier
authors.initials: X
authors.lastname: Duportet
authors.affiliation: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, New York, USA.
authors.forename: Vincent A
authors.initials: VA
authors.lastname: Fischetti
authors.affiliation: Laboratory of Bacteriology, The Rockefeller University, New York, New York, USA.
authors.forename: Luciano A
authors.initials: LA
authors.lastname: Marraffini
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000008:administration & dosage* / Q000138:chemical synthesis; D001435:Bacteriophages; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D004352:Drug Resistance, Microbial / Q000235:genetics*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects; D014774:Virulence / Q000187:drug effects / Q000235:genetics
chemical_list: D000890:Anti-Infective Agents