TABLE 1.
Use of Cas9 nickase instead of wild-type Cas9 for genome editing in C. cellulolyticum
| Plasmid | Component | Cell growth by resistance typea: |
|
|---|---|---|---|
| TMPr | 5-FOAr | ||
| pCas9 | Cas9 | Y | Ν |
| pGRNA-pyrF | gRNA | Y | Ν |
| pCas9-pyrF | Cas9 + gRNA | Ν | Ν |
| pCas9n-pyrF | Cas9n + gRNA | Y | Ν |
| pCas9-pyrF-donor | Cas9 + gRNA + donor template | Ν | Ν |
| pCas9n-pyrF-donor | Cas9n + gRNA + donor template | Y | Y |
a
TMPr, thiamphenicol resistant; 5-FOAr, 5-fluoroorotic acid resistant; Y, cell growth; Ν, no cell growth. Growth profiles are shown in Fig. S2 in the supplemental material.