TABLE 1.
The application of CRISPR/Cas9 and derived technologies in the biosynthesis of PNPs.
| Species | Products | Method | Regulation strategy | Culture method | Titer | References |
| E. coli | β-carotene | CRISPR/Cas9 | Genomic modification | Fed-batch (5 L) | 2.0 g/L | Li et al., 2015 |
| S. cerevisiae | Mevalonate | CRISPR/Cas9 | Multi-gene disruption | – | 1.5 mg/L (41-fold) | Jakounas et al., 2015 |
| Yarrowia lipolytica | Lycopene | CRISPR/Cas9 | Multi-gene integration | Shake flask (250 mL) | 8.6-fold | Schwartz et al., 2017 |
| S. cerevisiae | Valencene | CRISPR/Cas9 | Knock-out or down-regulation | Fed-batch (3 L) | 539.3 mg/L (160-fold) | Chen et al., 2019 |
| S. cerevisiae | guaia-6,10 (14)-diene | CRISPR/Cas9 | Gene integration | Fed-batch (5 L) | 0.8 g/L | Siemon et al., 2020 |
| Candida tropicalis | β-carotene | CRISPR/Cas9 | Multi-gene deletion or mutation | – | 0.23 mg/g DCW | Zhang L. et al., 2020 |
| Yarrowia lipolytica | β-carotene | CRISPR/Cas9 | Gene integration | Fed-batch (5 L) | 4.5 g/L | Zhang X.K. et al., 2020 |
| S. cerevisiae | β-carotene | CrEdit | Multi-loci gene integration | – | 12.7 mg/L | Ronda et al., 2015 |
| S. cerevisiae | Patchoulol | Cas-3P | Multiplexed and sequential editing | Shake flask (250 mL) | 20 mg/L | Li et al., 2020 |
| S. cerevisiae | Taxadiene | Cas9-based toolkit | Including Cas9-sgRNA plasmids, promoters, protein tags | Test tube (5 mL) | 20 mg/L (25-fold) | Apel et al., 2017 |
| E. coli | Naringenin | CRISPRi | Multi-gene repression | Shake flask | 421.6 mg/L (7.4-fold) | Wu et al., 2015 |
| E. coli | Lycopene (-)-α-bisabolol | CRISPRi | Regulatable repression | Shake flask | 50.6 mg/L 8.8-fold | Kim et al., 2016 |
| E. coli | Peonidin 3-O-glucoside | CRISPRi | Silence expression | Shake flask (125 mL) | 56 mg/L (2-fold) | Cress et al., 2017 |
| E. coli | Pinosylvin | CRISPRi | Multi-gene repression | Shake flask (500 mL) | 281 mg/L | Wu et al., 2017a |
| E. coli | Resveratrol | CRISPRi | Multi-gene down-regulation | Shake flask | 304.5 mg/L | Wu et al., 2017b |
| S. cerevisiae | α-amyrin | CRISPRi | Down-regulation | – | 11.97 mg/L | Yu et al., 2018 |
| S. cerevisiae | β-amyrin | CRISPRi | Multi-gene one-step down-regulation | Fed-batch (2.5 L) | 156.7 mg/L(1.44-fold) | Ni et al., 2019 |
| S. cerevisiae | β-carotene | CRISPR-AID | Synchronous up/down-regulation, deletion | – | 3-fold | Lian et al., 2017 |
| Ogataea polymorpha | Resveratrol | CMGE | Multi-gene knock-out, integration | Shake flask (500 mL) | 97.23 mg/L (20.73-fold) | Wang L. et al., 2018 |