TABLE 1

Primers, plasmids, and strains used in this study

Primer, plasmid, or strainSequence or descriptionaSource (reference)
Primers
    gRNA-uAATGAATTCTGGGCAAAGTAgttttragagctagaaatagcaagThis study
    gRNA-dTACTTTGCCCAGAATTCATTgatcatttatctttcactgcggaThis study
    Donor1-ucatccggagaacgaagcttctattaagacgatacagaatgaattctgAgcaaagtacggThis study
    Donor1-dttcaactttttcaaaatcataacgagtgaagaaagtacggccgtactttgcTcagaattThis study
    Donor2-ucatccggagaacgaagcttctattaagacgatacagaatgaattctgGgcaaagtacggThis study
    Donor2-dttcaactttttcaaaatcataacgagtgaagaaagtacggccgtactttgcCcagaattThis study
    Donor3-ucatccggagaacgaagcttctattaagacgatacagaatgaattctCagcaaagtacggThis study
    Donor3-dttcaactttttcaaaatcataacgagtgaagaaagtacggccgtactttgctGagaattThis study
    PGM2-ucatggaagctattccagagThis study
    PGM2-daccgttggttcttcagttccThis study
    PGM2-Seq-uccagcatggtcttctgtctacThis study
    PGM2-Seq-dgtacctgacaatcttagaacThis study
Plasmids
    Cas9-NATp414-TEF1p-Cas9-CYC1t-NAT1Addgene 64329 (50)
    p42K-gPGM2pRS42K carrying guide RNA for PGM2 deletionThis study
Strains
    S. boulardiiS. boulardii (ATCC-MYA-796)ATCC (38)
    S. cerevisiaeS. cerevisiae BY4742 (MATα his3Δ1 leu2Δ0 lys2Δ0 ura3Δ0)EUROSCARF (48)
    S. cerevisiae Δpgm2S. cerevisiae BY4742 with PGM2 deletionEUROSCARF (48)
    S. boulardii Δpgm2S. boulardii (ATCC-MYA-796) with PGM2 deletionThis study
    S. boulardii SBe1Isolated from serial subcultures of S. boulardii on galactoseThis study
    S. cerevisiae PGM2-TGAS. cerevisiae with mutated PGM2 (G1278A); stop codonThis study
    S. boulardii PGM2-TGGS. boulardii with mutated PGM2 (A1278G); mimic S. cerevisiaeThis study
    S. boulardii PGM2-TCAS. boulardii with mutated PGM2 (G1277C); mimic SBe1This study
  • a The capital letters are 20 base pairs of targeting sequences of guide RNA.