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Table 1 Summary of the genomic editing in M. thermophila using the CRISPR–Cas system

From: Upgrading of efficient and scalable CRISPR–Cas-mediated technology for genetic engineering in thermophilic fungus Myceliophthora thermophila

Host strainaTarget locusElements in co-transformationNo. of analyzed transformantsNo. of HR transformantsbHR efficiency (%)cEach gene disruption efficiency (%)
WTcre-1Donor-cre1-neo203150
WTcre-1Cas12a +donor-cre1-neo203150
WTcre-1crRNA +donor-cre1-neo202100
WTcre-1Cas12a + crRNA + donor-cre1-neo20189060
WTcre-1, res-1, gh1-1Donor DNA of cre-1, res-1 and gh1-123000
0
0
WTcre-1, res-1, gh1-1Cas12a + pooled three sets of crRNA + donor DNA of cre-1, res-1 and gh1-12084035
25
25
WTcre-1, res-1, gh1-1Cas12a + array1 + donor DNA of cre-1, res-1 and gh1-12273241
32
27
WTcre-1, res-1, gh1-1Cas9 + three sets of sgRNA + donor DNA of cre-1, res-1 and gh1-12393939
35
39
M3neo,alp-1, rca-1, hcr-1Donor DNA of neo, alp-1, rca-1 and hcr123000
0
0
0
M3neo,alp-1, rca-1, hcr -1Cas12a + array2 + donor DNA of neo, alp-1, rca-1 and hcr-12352226
30
13
22
M3neo, alp-1, rca-1, hcr-1Cas9 + four sets of sgRNA + donor DNA of neo, alp-1, rca-1 and hcr-12352235
30
13
30
M7bar, ap-3, prk-6Donor DNA of bar, ap-3 and prk-622000
0
0
M7bar, ap-3, prk-6Cas12a + array3 + donor DNA of bar, ap-3 and prk-62294132
27
23
M7Bar, ap-3, prk-6Cas9 + three sets of sgRNA + donor DNA of bar, ap-3 and prk-62183833
29
19
  1. aWT, wild-type strain; M3, triple-mutant Δcre1Δres1Δgh1-1; M7, septuple mutant ∆cre1Δres1Δgh1-1ΔneoΔalp1Δrca1::xyr1Δhcr1
  2. bHR, homologous recombination
  3. cHR efficiency, HR frequency of gene replacement (including both homokaryons and heterokaryon)