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. 2023 Nov 13;9(12):2494–2503. doi: 10.1021/acsinfecdis.3c00342

Figure 2.

Figure 2

Potency of CRISPR-Cas antifungals and potential rise of antifungal resistance. (A) Slower cell growth on solid media did not induce antifungal resistance. All selected strains carrying the top performing CRISPR-Cas antifungals exhibited loss of growth on galactose (Cas9 “on”) plate. (B) Faster cell growth on liquid media escaped CRISPR-Cas antifungal treatment. Three strains (Vip54, Vip57, and Vip59) exhibited a normal growth phenotype as compared to the control strain (Vip46*); nine strains (Vip47, Vip48, Vip55, Vip56, Vip69, Vip73, Vip74, Vip81, and Vip90) showed a growth lag but started to enter exponential growth after 48-h outgrowth; and the remaining five strains (Vip52, Vip60, Vip76, Vip93, and Vip94) exhibited growth arrest. (C) Comparison of potency between the top CRISPR-Cas antifungals and antibiotic Geneticin (G418). The percentage of viable cells for each strain is presented above each bar. Cell growth on glucose served as a negative control with inactive CRISPR-Cas systems, while cell characterization without a carbon source served as a positive control. In panels B and C, each value is a mean ± 1 standard deviation (n ≥ 2).