TABLE 1

Transformation efficiencies of a transient CRISPR-Cas9 system targeting ADE2 in C. lusitaniae haploid and diploid strains

CRISPR componentMean (SD) for strain type:
DiploidcHaploide
Deletion constructaCas9bsgRNAbNo. of NATr colonies% ade2dNo. of NATr colonies% ade2d
LongClusClus520 (107)7.9 (3.5)801 (495)36 (2)
CalbCalb211 (248)0281 (244)17.0 (13)
Clus282 (359)0.6 (0.8)341 (220)21.0 (17)
Clus418 (285)0.0 (0.1)879 (225)3.1 (3.0)
Calb82 (112)0.2 (0.3)335 (108)14.0 (9)
228 (281)0.4 (0.7)231 (71)8.4 (9.7)
ShortClusClus14 (19)041 (40)2.3 (3.1)
CalbCalb8.0 (13.0)076 (68)0
Clus13 (14)026 (14)0
Clus9.3 (14.5)018 (14)0
Calb1.0 (1.7)037 (22)0
1.7 (2.9)022 (6)0
  • a Deletion constructs had long flanks (~1-kb homology both 5′ and 3′ to the C. lusitaniae ADE2 gene) or short flanks (~80-bp homologous flanks).

  • b “Clus” and “Calb” indicate that the Cas9 and single guide RNA (sgRNA) are under a C. lusitaniae or C. albicans promoter, respectively. The 20-bp protospacer in the sgRNA was different between the haploid and diploid strains. Dashes indicate that a component was not included in the transformation.

  • c Diploid strain, C. lusitaniae CAY5019 (n = 3 transformations).

  • d ADE2 deletion percentage, number of red colonies/total number of NATr colonies × 100.

  • e Haploid strain, C. lusitaniae RSY284 (n = 3 transformations).