FIG 1.

CRISPR-Cas9-free and -dependent transformation strategies used for strain construction. Derived from the C. albicans reference strain SC5314, the parental SN148 strain was sequentially transformed using CRISPR-Cas9-free (gray) or CRISPR-Cas9-dependent (orange) strategy in order to integrate (i) the BFP/GFP LOH reporter system, (ii) a unique barcode sequence, and (iii) the leucine marker rendering the strains prototrophic. Strains constructed using the CRISPR-Cas9-free strategy underwent four transformation steps, as opposed to two using the CRISPR-Cas9-dependent strategy. The barcode sequence (BC) and LEU2 auxotrophic marker are introduced at the RPS1 locus on Chr1. The BC and LEU2 marker are introduced at another locus for the strains carrying the LOH reporter system on Chr1 [in the intergenic sequence between the CDR3 and tG(GCC)2 loci], as indicated by the striped arrows.