Table 1 Genome editing applications of CRISPR–Cas9 in non-conventional yeasts
From: CRISPR-mediated genome editing in non-conventional yeasts for biotechnological applications
| Yeasts | Cas9 promoter | sgRNA promoter | Host strains | Target locus | NHEJ efficiency (positive colonies/total colonies) | HR efficiency (positive colonies/total colonies), donor length | References |
|---|---|---|---|---|---|---|---|
| S. stipiti | ENO1 | SNR52 | UC7 | ADE2 | 83%, (5/6) | [33] | |
| TRP1 | 100%, | ||||||
| UC7 ku70Δku80Δ | ADE2 | 71%, (5/7), 500 bp HA | [35] | ||||
| TRP1 | 64%, (14/22), 50 bp HA | ||||||
| UC7 | KU70 | 100%, (4/4) | |||||
| KU80 | 83%, (5/6) | ||||||
| K. phaffii | HTA1 | HTB1 | CBS7435 | GUT1 | 94%, (79/84) | [39] | |
| AOX1 | 95% | ||||||
| GUT1+AOX1 | 69% | ||||||
| MPP1 | 95% | ||||||
| TRM1 | 94% | ||||||
| MXR1 | 43% | ||||||
| OCH1 | 51% | ||||||
| CBS7435 ku70Δ | GUT1 | 91%, 1000 bp HA | [41] | ||||
| O. polymorpha | TDH3 | tRAN CUG | BY4330 | ADE12 | 45%, (48/106) | 47%, (186/394), 60 bp HA | [42] |
| AaTEF1 | ScTDH3 | CBS4732 | ADE2 | 9% | [43] | ||
| ScTEF1 | ScSNR52 | LEU2 | 58% | [45] | |||
| URA3 | 65% | ||||||
| ADE2 | 62%, 1000 bp HA | ||||||
| URA3+HIS3+LEU2 | 24%, 1000 bp HA | ||||||
| O. thermomethanolica | AOX | AOX | TBRC656 | HAC1 | 63%, (15/24) | [47] | |
| MAL1 | 97%, (126/130) | ||||||
| MAL2 | 93%, (37/40) | ||||||
| K. lactis | FBA1 | SNR52 | ATCC8585 Gal80Δ- Ku80Δ | DIT1+ADH1+NDT80 | 2%, 500 bp HA | [49] | |
| K. marxianus | ScPDC1 | ScSNR52 | NBR1777 Nej1Δ | URA3 | 84%, (18/19), 120 bp HA | [52] | |
| NBR1777 Dnl4Δ | 100%, (20/20), 120 bp HA | ||||||
| NBR1777 Nej1Δ | Sed1 | 84%, (6/7), 50 bp HA | |||||
| NBR1777 Dnl4Δ | 92%, (46/50), 50 bp HA | ||||||
| TEF1 | RPR1-tRNA Gly | CBS6556 | XYL2 | 66%, (68/90) | [53] | ||
| Y. lipolytica | UAS1B8-TEF1 |
SCR1’-
tRNA Gly |
PO1f (ATCC MYA-2613) |
KU70
PEX10 MFE1 |
100% 54%, (16/30) 90%, (27/30) | [55] | |
| PO1f Ku70Δ |
PEX10
MFE1 |
86%, (30/35), 1000 bp HA 100%, (7/7), | |||||
| PO1f |
MFE1
AXP XPR2 XDH XYR | \( \left. \begin{aligned} 69\% , \hfill \\ 62\% , \hfill \\ 48\% , \hfill \\ 53\% , \hfill \\ 52\% \hfill \\ \end{aligned} \right\}1000 \; {\text{bp}}\;{\text{HA}} + {\text{GFP}} \) | [59] | ||||
| TEFin | TEFin | PO1f | TRP1 | 13%, (10/76) | [55] | ||
|
PO1f Ku70Δ PO1f Ku70ΔKu80Δ | \( \left. \begin{aligned} 1 1\% , \, \left( { 1 1/ 9 8} \right) \hfill \\ 100\% , \, \left( { 1 9/ 1 9} \right) \hfill \\ 100\% , \, \left( { 1 5/ 1 5} \right) \hfill \\ \end{aligned} \right\} 500{\text{ bp HA}} \) | [58] | |||||
| PO1f | PEX10 | \( \left. \begin{aligned} 6 1\% , \, \left( { 1 7/ 2 8} \right) \hfill \\ 3 3\% , \, \left( { 4/ 1 2} \right) \hfill \\ \end{aligned} \right\} 500{\text{ bp HA}}\; {\text{in plasmid}} \) | |||||
| TRP1+PEX10 | 37%, (15/40) | ||||||
| TRP1+PEX10+GUT2 | 20%, (7/35) |