Higher
cell inoculation is more prone to CRISPR-Cas antifungal
resistance. (A–F) Effect of cell inoculation concentrations
on the potency of CRISPR-Cas antifungals. Five strains (Vip52, Vip60,
Vip76, Vip93, and Vip94) harboring the most effective antifungal designs
and one control strain (Vip46*) were investigated with an initial
OD of either 0.05 or 0.2. Higher concentration of cells at the outset
of the experiment overwhelmed the population by decreasing lag time,
thereby increasing both antibiotic and CRISPR-Cas antifungal resistance.
(G) Viable cell count at 36-h outgrowth confirmed the increase in
viable cells in the high inoculation scenario. In panels A–G,
each value is a mean ±1 standard deviation (n ≥ 2).