Extended Data Fig. 7: Genomic targeting of P. aeruginosa PAO1 with all-in-one vector pCas3ch.
From: A compact Cascade–Cas3 system for targeted genome engineering
a, Map of the I-C CRISPR-Cas all-in-one plasmid pCas3cRh carrying I-C crRNA and genes cas3, cas5, cas8, and cas7 under the control of the rhamnose-inducible rhaSR-PrhaBAD system. b, Growth curve of PAO1 transformed with the pCas3cRh vector expressing a self-targeting crRNA targeting phzM (Ind.). An empty vector (E.V.) and a non-induced phzM targeting strain (N.I.) were used as controls. Mean OD values measured at 600 nm are shown for six biological replicates each. c, Deletion efficiencies for WT PAO1 using the all-in-one vector pCas3cRh carrying all necessary components of the I-C CRISPR-Cas system. Values are averages of three replicates where 12 individual colonies were analyzed using site-specific PCR. Error bars show standard deviations. d, Transformation efficiencies with self-targeting pCas3cRh vectors expressing crRNAs for phzM or XNES 2 compared to a non-targeting control (green bar) in PAO1. Values are means of 3 replicates each, error bars represent SD values.
