Table 2 CRISPR/Cas9-mediated homologous recombination (HR) efficiency of simultaneous disruption of one to four gene loci
Host strain | Target locus | Elements in co-transformation | No. of analyzed transformants | No. of HR transformants | HR efficiency (%) |
|---|---|---|---|---|---|
ATCC 42464 | cre-1 | Donor-cre1 | 20 | 4 | 20 |
ATCC 42464 | cre-1 | Cas9Â +Â donor-cre1 | 20 | 3 | 15 |
ATCC 42464 | cre-1 | cre1-sgRNAÂ +Â donor-cre1 | 20 | 3 | 15 |
ATCC 42464 | cre-1 | Cas9Â +Â cre1-sgRNAÂ +Â donor-cre1 | 20 | 19 | 95 |
CBS 203.75 | cre-1 | Donor-cre1 | 20 | 4 | 20 |
CBS 203.75 | cre-1 | Cas9Â +Â donor-cre1 | 20 | 3 | 15 |
CBS 203.75 | cre-1 | cre1-sgRNAÂ +Â donor-cre1 | 20 | 4 | 20 |
CBS 203.75 | cre-1 | Cas9Â +Â cre1-sgRNAÂ +Â donor-cre1 | 20 | 18 | 90 |
M2 strain | res-1, gh1-1 | Two sets of sgRNA and donor DNA of res-1 and gh1-1 | 23 | 15 | 65 |
M2 strain | cre-1, gh1-1 | Two sets of sgRNA and donor DNA of cre-1 and gh1-1 | 23 | 14 | 61 |
M2 strain | cre-1, res-1 | Two sets of sgRNA and donor DNA of cre-1 and res-1 | 23 | 16 | 70 |
M2 strain | cre-1, res-1, gh1-1 | Three sets of sgRNA and donor DNA of cre-1, res-1 and gh1-1 | 23 | 7 | 30 |
M2 strain | cre-1, res-1, gh1-1, alp-1 | Four sets of sgRNA and donor DNA of cre-1, res-1, gh1-1 and alp-1 | 23 | 5 | 22 |