Table 1 Application status of CRISPR/Cas system in microalgae
Microalgae species | Microalgae subtypes | Editing method | Transformation method | Editing efficiency | Experimental results | References |
|---|---|---|---|---|---|---|
C. Reinhardtii | C. reinhardtii CC-503 | One plasmid-driven CRISPR/Cas9 system (C. reinhardtii codon-optimized Cas9) | Electroporation | 46.7% | First successful transient expression of Cas9 and sgRNA in C. reinhardtii | [23] |
C. reinhardtii CC-124 | CRISPR/Cas9 RNPs | Electroporation | 0.17–40% | Mutagenesis of MAA7, CpSRP43 and ChlM gene | [24] | |
/ | CRISPR/Cas9 RNPs | Electroporation | 0.56% | CpFTSY and ZEP gene knockout, increases photosynthetic productivity | [25] | |
C. reinhardtii CC-400 | Two plasmid-driven CRISPRi/dCas9-KRAB system | Glass beads | 94% | Downregulating the expression level of CrPEPC1 gene to increase lipid synthesis | [26] | |
C. reinhardtii CC-4349 | CRISPR/Cas9 RNPs | Natural transformation | / | Knock-out the zeaxanthin epoxidase gene to stop the formation of lutein | [27] | |
/ | CRISPR/Cas12a RNPs | Electroporation | / | Exploring the mechanisms of single-strand templated DNA repair at CRISPR/Cas12a-induced DSBs in C. reinhardtii | [28] | |
C. Reinhardtii CC-4349, CC-124, and CC-503 | CRISPR/Cas9 RNPs | Electroporation | 30% | Knock-in antibiotics resistance gene and YFP | [29] | |
Synechococcus elongatus | Synechococcus elongatus PCC 7942 | One plasmid-driven CRISPRi/dCas9 system | Natural transformation | 99% | The glgc gene was downregulated to increase succinate titer level | [30] |
Synechococcus elongatus UTEX 2973 | Two plasmid-driven CRISPR/Cas9 system | Natural transformation | 30–100% | Knock-out the nblA gene | [31] | |
Synechococcus sp. | Synechococcus sp. PCC 7002 | Genomic integration CRISPRi/dCas9 system | Natural transformation | 30–90% | Increasing central carbon flux by reducing carboxysome expression level | [32] |
Synechococcus sp. PCC7942 | One plasmid-driven CRISPR/Cas9 system | Natural transformation | 23–57% | Improvement of succinate synthesis using glgc gene knock-out and gltA/ppc gene knock-in | [33] | |
Synechocystis sp. PCC 6803 | One plasmid-driven CRISPRi/dCas9 system | Natural transformation | 50–95% | Prevent the synthesis of carbon storage compounds | [34] | |
N. oceanica | N. oceanica CCMP1779 | One plasmid-driven CRISPR/Cas9 system | Electroporation | 45–90% | Generating non-transgenic marker-free nitrate reductase knock-out lines | [35] |
N. oceanica IMET1 | CRISPR/Cas9 RNPs | Electroporation | 93% | FnCas12a as the best performer for genome editing in Nannochloropsis oceanica IMET1 | [36] | |
Chlorella | / | One plasmid-driven CRISPR/Cas9 system | Electroporation | 67% | Enhancing lipid accumulation | [37] |
C. vulgaris UTEX395 | CRISPR/Cas9 RNPs | Electroporation | / | Successful genome editing in C. vulgaris UTEX395 with CRISPR/Cas9 system | [38] | |
T. pseudonana | / | One plasmid-driven CRISPR/Cas9 system | Biolistic bombardment | 61.5% | Two sgRNAs are used to induce a precise deletion in the urease gene of T. pseudonana | [39] |
P. tricornutum | / | One plasmid-driven CRISPR/Cas9 system (C. reinhardtii codon-optimized Cas9) | Biolistic bombardment | 25–63% | Mutagenesis of the CpSRP54 gene to increase the sensitivity to high intensity light | [40] |