TABLE 1

Strains and plasmids used in this study

Strain or plasmidDescriptionaSource or reference
Strains
    E. coliTrans1-T1Clone host strain, F ϕ80(lacZ)ΔM15 ΔlacX74 hsdR (rK mK+) ΔrecA1398 endA1 tonATransGen
    S. aureus
        RN42208325-4, restriction-negative strainLaboratory stock
        NCTC 8325Wild typeLaboratory stock
        N315HA-MRSALaboratory stock
        RSD0RN4220 carrying plasmid pSD1This study
        RSD1RN4220 carrying plasmid pSD1-murEThis study
        RSD2RN4220 carrying plasmid pSD1-rpsCThis study
        RSD3RN4220 carrying plasmid pSD1-walRThis study
        NSD0NCTC 8325 carrying plasmid pSD1This study
        NSD1NCTC 8325 carrying plasmid pSD1-hla1This study
        NSD2NCTC 8325 carrying plasmid pSD1-hla2This study
        NSD3NCTC 8325 carrying plasmid pSD1-hla3This study
        NSD4NCTC 8325 carrying plasmid pSD1-hla4This study
        NSD5NCTC 8325 carrying plasmid pSD1-sarT1This study
        NSD6NCTC 8325 carrying plasmid pSD1-sarT2This study
        NSD7NCTC 8325 carrying plasmid pSD1-spa1This study
        NSD8NCTC 8325 carrying plasmid pSD1-hla1&2This study
        NSD9NCTC 8325 carrying plasmid pSD1-spa1hla2This study
        NSD10NCTC 8325 carrying plasmid pSD1-era1This study
        NSD11NCTC 8325 carrying plasmid pSD1-era2This study
        NSD12NCTC 8325 carrying plasmid pSD1-era3This study
        NSD13NCTC 8325 carrying plasmid pSD1-era4This study
        MSD0N315 carrying plasmid pSD1This study
        MSD1N315 carrying plasmid pSD1-mecA1This study
        MSD2N315 carrying plasmid pSD1-ccrAB1This study
        MSD3N315 carrying plasmid pSD1-ccrAB2This study
        MSD4N315 carrying plasmid pSD1-ccrAB3This study
        MSD5N315 carrying plasmid pSD1-ccrAB4This study
Plasmids
    pRMC2Shuttle vector, ATc inducible, Apr Cmr 22
    pSD1pRMC2 derivative with dCas9 and sgRNA expression cassetteThis study
    pSD1-hla1pSD1 with sgRNA1 targeting hlaThis study
    pSD1-hla2pSD1 with sgRNA2 targeting hlaThis study
    pSD1-hla3pSD1 with sgRNA3 targeting hlaThis study
    pSD1-hla4pSD1 with sgRNA4 targeting hlaThis study
    pSD1-hla1&2pSD1 with sgRNA1 and sgRNA2 targeting hlaThis study
    pSD1-sarT1pSD1 with sgRNA1 targeting sarTThis study
    pSD1-sarT2pSD1 with sgRNA2 targeting sarTThis study
    pSD1-spa1pSD1 with sgRNA1 targeting spaThis study
    pSD1-spa1hla2pSD1 with sgRNA1 targeting spa and sgRNA2 targeting hlaThis study
    pSD1-mecA1pSD1 with sgRNA1 targeting mecAThis study
    pSD1-ccrAB1pSD1 with sgRNA1 targeting ccrABThis study
    pSD1-ccrAB2pSD1 with sgRNA2 targeting ccrABThis study
    pSD1-ccrAB3pSD1 with sgRNA3 targeting ccrABThis study
    pSD1-ccrAB4pSD1 with sgRNA4 targeting ccrABThis study
    pSD1-era1pSD1 with sgRNA1 targeting operon containing eraThis study
    pSD1-era2pSD1 with sgRNA2 targeting operon containing eraThis study
    pSD1-era3pSD1 with sgRNA3 targeting operon containing eraThis study
    pSD1-era4pSD1 with sgRNA4 targeting operon containing eraThis study
    pSD1-murEpSD1 with sgRNA targeting murEThis study
    pSD1-rpsCpSD1 with sgRNA targeting rpsCThis study
    pSD1-walRpSD1 with sgRNA targeting walRThis study
  • a Abbreviations: Apr, ampicillin-resistant; Cmr, chloramphenicol-resistant.