Table 1 The genome editing efficiency of CRISPR/Cas9 system for H. insolens in this study
Host strain | Target gene | DNA fragments used for transformation | Number of target gene disruption transformants | Number of analyzed transformants | Number of all transformants | Gene editing efficiency (%) | |
|---|---|---|---|---|---|---|---|
Y1 | pks | Cas9 + sgRNA | Cas9 + 5SsgRNA-pks1 | 14 | 345 | 557 | 4.1 |
Cas9 + tRNAsgRNA-pks1 | 35 | 375 | 594 | 9.3 | |||
Cas9 + 5SsgRNA-pks1-HDV | 26 | 371 | 590 | 7.0 | |||
Cas9 + 5SHDVsgRNA-pks1 | 79 | 336 | 527 | 23.5 | |||
Cas9 + 5StRNAsgRNA-pks1 | 186 | 322 | 534 | 57.8 | |||
Cas9 + 5SHDVsgRNA-pks2 | 71 | 317 | 536 | 22.4 | |||
Cas9 + 5StRNAsgRNA-pks2 | 194 | 328 | 549 | 59.1 | |||
Cas9 + 5SHDVsgRNA-pks3 | 60 | 281 | 492 | 21.4 | |||
Cas9 + 5StRNAsgRNA-pks3 | 208 | 369 | 611 | 56.4 | |||
Donor DNA + sgRNA | Donor-pks + 5SsgRNA-pks1-HDV | 11 | 363 | 586 | 3.0 | ||
Donor-pks + 5SHDVsgRNA-pks1 | 11 | 351 | 556 | 3.1 | |||
Donor-pks + 5StRNAsgRNA-pks1 | 17 | 394 | 618 | 4.3 | |||
Donor DNA + Cas9 | Donor-pks + Cas9 | 18 | 383 | 617 | 4.7 | ||
Cas9 + donor DNA + sgRNA | Cas9 + donor-pks + 5SHDVsgRNA-pks1 | 91 | 342 | 562 | 26.6 | ||
Cas9 + donor-pks + 5StRNAsgRNA-pks1 | 204 | 363 | 593 | 56.2 | |||
Donor DNA | Donor-pks | 25 | 400 | 625 | 6.3 | ||
Y1/Δku70 | pks | Donor DNA | Donor-pks | 8 | 73 | 143 | 11.0 |
Cas9 + donor DNA + sgRNA | Cas9 + donor-pks + 5SHDVsgRNA-pks1 | 10 | 39 | 92 | 25.6 | ||
Cas9 + donor-pks + 5StRNAsgRNA-pks1 | 37 | 44 | 85 | 84.1 | |||
xyr1 | Cas9 + donor-xyr1 + 5SHDVsgRNA-xyr1 | 6 | 23 | 76 | 26.1 | ||
Cas9 + donor-xyr1 + 5StRNAsgRNA-xyr1 | 18 | 23 | 81 | 78.3 | |||
pks + xyr1 | Cas9 + donor-pks + donor-xyr1 + 5StRNA sgRNA-pks1 + 5StRNAsgRNA-xyr1 | 4 | 23 | 38 | 17.4 | ||