Table 1.
Nuclease activity of CRISPR protein orthologs in yeast
| Nuclease | gRNA | Protein-NLS | NLS-Protein-NLS |
|---|---|---|---|
| SpCas9 | Sg10 | – | ~ 80% |
| NmCas9 | Sg12 | – | 0 |
| Sg13 | – | 0 | |
| Sg29 | – | 0 | |
| Sg32 | – | 0 | |
| St1Cas9 | Sg14 | 0 | ~ 62% |
| Sg15 | 0 | 0 | |
| Sg30 | 0 | ~ 2.4% | |
| Sg64 | 0 | ~ 72% | |
| SaCas9 | Sg31 | ~ 50% | ~ 46% |
| Sg93 | ~ 27% | ~ 30% | |
| Sg94 | ~ 4.6% | ~ 5.2% | |
| Sg95 | ~ 77% | ~ 84% | |
| AsCpf1 | Sg68 | 0 | ~ 0.2% |
| LbCpf1 | Sg69 | 0 | ~ 59% |
| Sg122 | 0 | ~ 92% | |
| Sg123 | 0 | ~ 55% | |
| Sg124 | 0 | ~ 0.3% |
Nuclease activity was evaluated by co-transforming 500 ng CRISPR protein plasmid, 500 ng gRNA plasmid, and 500 ng linear DNA donor for the deletion of whole ADE2 coding sequences. The results represent an average of biological triplicates