TABLE 3.
Efficiency of target gene insertion and replacement in the G. lucidum Cas9 strain and the ku70 mutanta
| Strain | Mutation of ura3 | Expt. 1 | Expt. 2 | Expt. 3 | Efficiency (%) |
|---|---|---|---|---|---|
| Cas9 | Insertion | 1/20 | 1/23 | 0/17 | 3.3 |
| ku70 mutant | Insertion | 10/10 | 8/8 | 8/9 | 96.3 |
| Cas9 | Replacement | 0/13 | 0/10 | 0/11 | 0 |
| ku70 mutant | Replacement | 7/8 | 9/9 | 11/12 | 93.1 |
a
Targeted gene insertions were screened for in 2 of 60 5-FOA-resistant G. lucidum Cas9 transformants produced and in 26 of 27 5-FOA-resistant ku70 mutant transformants produced, in three independent experiments. Targeted gene replacement was identified in 0 of 34 5-FOA-resistant G. lucidum Cas9 transformants produced and in 27 of 29 5-FOA-resistant ku70 mutant transformants produced, in three independent experiments.