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).