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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 straina Target locus Elements in co-transformation No. of analyzed transformants No. of HR transformantsb HR efficiency (%)c Each gene disruption efficiency (%)
WT cre-1 Donor-cre1-neo 20 3 15 0
WT cre-1 Cas12a +donor-cre1-neo 20 3 15 0
WT cre-1 crRNA +donor-cre1-neo 20 2 10 0
WT cre-1 Cas12a + crRNA + donor-cre1-neo 20 18 90 60
WT cre-1, res-1, gh1-1 Donor DNA of cre-1, res-1 and gh1-1 23 0 0 0
0
0
WT cre-1, res-1, gh1-1 Cas12a + pooled three sets of crRNA + donor DNA of cre-1, res-1 and gh1-1 20 8 40 35
25
25
WT cre-1, res-1, gh1-1 Cas12a + array1 + donor DNA of cre-1, res-1 and gh1-1 22 7 32 41
32
27
WT cre-1, res-1, gh1-1 Cas9 + three sets of sgRNA + donor DNA of cre-1, res-1 and gh1-1 23 9 39 39
35
39
M3 neo,alp-1, rca-1, hcr-1 Donor DNA of neo, alp-1, rca-1 and hcr1 23 0 0 0
0
0
0
M3 neo,alp-1, rca-1, hcr -1 Cas12a + array2 + donor DNA of neo, alp-1, rca-1 and hcr-1 23 5 22 26
30
13
22
M3 neo, alp-1, rca-1, hcr-1 Cas9 + four sets of sgRNA + donor DNA of neo, alp-1, rca-1 and hcr-1 23 5 22 35
30
13
30
M7 bar, ap-3, prk-6 Donor DNA of bar, ap-3 and prk-6 22 0 0 0
0
0
M7 bar, ap-3, prk-6 Cas12a + array3 + donor DNA of bar, ap-3 and prk-6 22 9 41 32
27
23
M7 Bar, ap-3, prk-6 Cas9 + three sets of sgRNA + donor DNA of bar, ap-3 and prk-6 21 8 38 33
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)