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Table 1 Comparison on CRISPR/Cas9 system for A. niger

From: Heterologous and endogenous U6 snRNA promoters enable CRISPR/Cas9 mediated genome editing in Aspergillus niger

Cas9 expression sgRNA expression Targeted gene Donor DNA (SM/homology arm size, bp) Gene editing efficiency Notes References
Promoter Terminator Promoter Terminator
Ptef1 Ttef1 PgpdA TtrpC albA Some Requiring to add HH and HDV for processing sgRNA [10]
PcoxA Ttef1 PmbfA TtrpC pyrG Obtaining 25 colonies on MM with 5′-FOA Requiring to add self-cleaving ribozymes for processing sgRNA [11]
MttA pyrG/690 and 834b 100% (7/7)
Ptef1 Ttef1 In vitro synthesis 1090836 pyrG/1500 28% (11/40) 3% (1/30)a [12]
1117792 100% (8/8) 43% (13/30)a
1141260 100% (8/8) 0% (0/30)a
1121140 38% (3/8) 2% (1/60)a
1146483 88% (7/8)  
1170646 63% (5/8)  
PglaA TglaA PhU6 Ploy(T)6 albA 15% (4/27) Without any selection pressure for targeted gene editing This study
PyU6    20% (1/5)
PanU6    23% (3/13)
PanU6   albA hph/40 36% (5/14) 79% (11/14)c
  1. aThe gene deletion efficiency without CRISPR/Cas9 system
  2. bThis donor DNA for gene integration was flanked by a 5′ flanking sequence of 690 bp homologous to the promoter region of the pyrG gene, while the 3′ flanking sequence was a mutated and truncated pyrG CDS of 834 bp (pyrGm2, trunc)
  3. cAfter co-transformation of donor DNA MHi-albA-hph, sgRNA3.1 and pCas9, the outgrown albino colonies with albA disruption reached up to 79% (11/14). In some albino colonies, some unexpected base pair errors were mediated by NHEJ at the 5′-junction or 3′-junction of DBSs. Therefore, the precise gene integration was only 36% (5/14)