Table 3. Simultaneous multiple chromosome splitting by CRISPR-PCS.
| Splitting pointa | CRISPR -PCS | Length of homology sequence | No. of transformants | No. of transformants subjected to karyotype analysis | No. of transformants with expected splitting |
|---|---|---|---|---|---|
| C4-P3 (URA3) C15-P1 (CgHIS3) C16-P1(CgLEU2) | + | 50 | 206 | 10 | 4 |
| − | 50 | 0 | 0 | 0 | |
| C12-P1 (KanMX) C15-P1 (CgHIS3) C16-P1(CgLEU2) | + | 50 | 414 | 38b | 6b |
| − | 50 | 0 | 0 | 0 | |
| C4-P1 (CgHIS3) C4-P2(CgLEU2) C4-P3 (URA3) | + | 50 | 88 | 10 | 2 |
| − | 50 | 0 | 0 | 0 | |
| C4-P3 (URA3) C12-P1(KanMX) C15-P1 (CgHIS3) C16-P1(CgLEU2) | + | 50 | 70 | 20 | 4 |
| − | 50 | 0 | 0 | 0 |
aOne of the two splitting modules for each splitting site contains a marker gene described in parentheses.
bConfirmation of expected splitting event was performed by colony direct PCR.