Strains not associated with the library are summarized in Supplementary file 8 . C. albicans transformations were performed using the standard lithium acetate transformation method (Noble and Johnson, 2005 (link)). The single and double homozygous mutant strains of C. albicans were constructed from an SN152 background using the transient CRISPR/Cas9 method (Min et al., 2016 (link)). Oligonucleotides and plasmids used to generate the mutant strains in this study are listed in Supplementary file 9 . Briefly, the ume6∆∆ mutant strain was generated by deleting one copy of UME6 with HIS1 cassette which was amplified from pFA-LHL plasmid (Dueñas-Santero et al., 2019 (link)) with primer pairs UME6.P1 and UME6.2. The second allele of UME6 was replaced with the ARG4 marker amplified from pFA-LAL (Min et al., 2016 (link)) plasmid with primer pairs UME6.P1 and UME6.P2 and by using sgRNA targeting individual alleles of UME6 gene.
The brg1∆∆ mutant strain was generated by amplifying ARG4 cassette from pSN69 plasmid (Noble and Johnson, 2005 (link)) with primer pairs BRG1.P1 and BRG1.P2 and by using sgRNA targeting two alleles of BRG1 gene. The rob1∆∆ homozygous strain was generated by amplifying HIS1 cassette from the pSN52 plasmid (Noble and Johnson, 2005 (link)) with primer pairs ROB1.P1 and ROB1.P2 by using sgRNA targeting two alleles of ROB1 gene. The resulting brg1∆∆ and rob1∆∆ mutants were further used to generate double homozygous brg1∆∆ nrg1∆∆ and rob1∆∆ nrg1∆∆. To do this, both copies of NRG1 knocked out by amplifying HIS1 or ARG4 cassette from the plasmid pFA-LHL or pFA-LAL, respectively, with primer pairs NRG1.P1 and NRG1.P2 and using sgRNA targeting two alleles of NRG1 gene. The efg1∆∆ mutant strain from Homann collection (Homann et al., 2009 (link)) was used to generate the double homozygous efg1∆∆ nrg1∆∆ mutant. To do this, pFA-LAL plasmid was used to amplify the ARG4 cassette using NRG1.P1 and NRG1.P2 primer pairs and using sgRNA targeting two alleles of NRG1 gene. The resultant transformants were selected on the SD plates lacking either histidine or arginine. The single or double homozygous integration of the deletion cassette was confirmed by standard PCR methods.
Fluorescently labeled strains were generated by using pENO1-NEON-NAT1 and pENO1-iRFP-NAT1 plasmids as previously described (Bergeron et al., 2017 (link); Seman et al., 2018 (link)) and the resultant transformants were selected on YPD containing 200 µg/ml nourseothricin (Werner Bioagents, Jena, Germany). The reference strain was tagged with green fluorescent protein (NEON) whereas all the TF mutants were tagged with iRFP.
The brg1∆∆ mutant strain was generated by amplifying ARG4 cassette from pSN69 plasmid (Noble and Johnson, 2005 (link)) with primer pairs BRG1.P1 and BRG1.P2 and by using sgRNA targeting two alleles of BRG1 gene. The rob1∆∆ homozygous strain was generated by amplifying HIS1 cassette from the pSN52 plasmid (Noble and Johnson, 2005 (link)) with primer pairs ROB1.P1 and ROB1.P2 by using sgRNA targeting two alleles of ROB1 gene. The resulting brg1∆∆ and rob1∆∆ mutants were further used to generate double homozygous brg1∆∆ nrg1∆∆ and rob1∆∆ nrg1∆∆. To do this, both copies of NRG1 knocked out by amplifying HIS1 or ARG4 cassette from the plasmid pFA-LHL or pFA-LAL, respectively, with primer pairs NRG1.P1 and NRG1.P2 and using sgRNA targeting two alleles of NRG1 gene. The efg1∆∆ mutant strain from Homann collection (Homann et al., 2009 (link)) was used to generate the double homozygous efg1∆∆ nrg1∆∆ mutant. To do this, pFA-LAL plasmid was used to amplify the ARG4 cassette using NRG1.P1 and NRG1.P2 primer pairs and using sgRNA targeting two alleles of NRG1 gene. The resultant transformants were selected on the SD plates lacking either histidine or arginine. The single or double homozygous integration of the deletion cassette was confirmed by standard PCR methods.
Fluorescently labeled strains were generated by using pENO1-NEON-NAT1 and pENO1-iRFP-NAT1 plasmids as previously described (Bergeron et al., 2017 (link); Seman et al., 2018 (link)) and the resultant transformants were selected on YPD containing 200 µg/ml nourseothricin (Werner Bioagents, Jena, Germany). The reference strain was tagged with green fluorescent protein (NEON) whereas all the TF mutants were tagged with iRFP.
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