Extended Data Figure 6: Cas9 fusion to ORF152 localizes to periphery of shell, but does not enable immune activity against ΦKZ.

a, Strain PAO1 was engineered to express Cas9 fused to ORF152 at either the N- or C-terminus, with single guide RNAs (sgRNAs) targeting the indicated phage. Plaque assays were conducted as in Fig. 1a, b, growth curves were conducted, monitoring the OD₆₀₀ of PAO1 cells infected with the indicated ΦKZ multiplicity of infection (MOI). c, Fluorescence microscopy of PAO1 fusion strains, immunostained for Cas9, in cells expressing ORF152-Cas9 or Cas9-ORF152. DAPI stain shows the phage DNA within the shell. d, Fluorescence microscopy of P. aeruginosa, immunostained for Cas9, in cells expressing ORF152-Cas9. DAPI stain shows the phage DNA within the shell. All plaque assays were replicated 4 times with similar results. Growth curve experiments were replicated three times with similar results. Microscopy was replicated twice with similar results.