FIG 2.

Acrs-mediated CRISPR-Cas inactivation requires a critical phage concentration. (A) PA14 was treated with DMS3_acrIF1/DMS3m_acrIF1 for 12 h (1 × 10⁷ PFU, MOI = 0.02) and infected with DMS3_acrIF1/DMS3 or DMS3m_acrIF1/DMS3m for 36 h (1 × 10⁷ PFU, MOI = 0.02). Growth kinetics upon phage infection were continuously monitored from the initial DMS3_acrIF1 challenge to 48 h. (B) Inhibitory rates (%) of DMS3_acrIF1 on PA14 were evaluated within 48 h after treatment with DMS3_acrIF1 for 12 h and were infected with DMS3 or DMS3_acrIF1 for 36 h (1 × 10⁷ PFU, MOI = 0.02). (C) DMS3_acrIF1 of different titers was used to infect PA14 (5 × 10⁸ CFU/mL), and the growth kinetics upon phage infection were calculated by measuring the OD₆₀₀ nm at 20 min intervals up to 48 h at 37°C with constant shaking. Data were presented as mean ± standard error of the mean (SEM), determined from biological triplicates and analyzed via a one-way ANOVA compared against a control group (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).