Table 1.
Decrease in polyunsaturated fatty acid by CRISPR/Cas9.
| Gene name | Technique | Promoter of Cas9 | Method | Phenotype | Oleic acid WT (%) | Oleic acid Mutant (%) | Mutation type | References |
|---|---|---|---|---|---|---|---|---|
|
AhFAD2A
AhFAD2B |
CRISPR/Cas9 | CamV 35S | Hairy root transformation | - | 36 ~ 67% | Not harvest the seeds | G448A (ahFAD2A), +1 bp, and G451T (ahFAD2B) | Yuan et al., 2019 |
| NtFAD2-2 | CRISPR/Cas9 | CamV 35S | Agrobacterium-mediated transformation | No side effect | ~12% | 79% | -1 and −5 bp | Tian et al., 2020a |
| AtFAD2 | Base editing | RPS5A (Arabidopsis) | Floral dipping | Resistance to salt stress | 18.5% | 57.9% | A295G, D298E | Park et al., 2021 |
| 64.7% | A295V, T296M | |||||||
| 30.6% | A295V | |||||||
| 29.6% | A295G | |||||||
| CRISPR/Cas9 | CamV 35S | Floral dipping | - | 16.2% | ~59.8% | +1 bp | Jiang et al., 2017 | |
| BnFAD2 | CRISPR/Cas9 | Ubiquitin4-2 (Petroselinum crispum) | Agrobacterium-mediated transformation | No difference | cv. Westar(74.6%) | BnFAD2_Aa (80%) | -4 bp | Okuzaki et al., 2018 |
| Ubiquitin (rice) | cv. J9707 (66.7%) |
BnaFAD2.A5 (73.1–82.3%) | −1bp, −1bp and S1, −2bp, −13bp, −80bp, +1bp, +1bp and +1bp, +1bp and −2bp, +1bp and −7bp | Huang et al., 2020 | ||||
| BnaFAD2.C5 (73–74%) | −3 and +1 bp | |||||||
| CsFAD2 | CRISPR/Cas9 | Ubiquitin4-2 (Petroselinum crispum) | Floral dipping | All CsFAD2 gene mutants → slow growth, twisted leaves, delayed bolting |
cv. Celine (9.8%) |
10–62% | 21 different mutant alleles | Morineau et al., 2017 |
| CamV 35S | - | cv. Suneson (15.9%) |
~54.7% | A lot of mutant alleles | Jiang et al., 2017 | |||
| EC1.2 | All CsFAD2 gene mutants → Stunted bushy phenotype, small, and bloomed late | cv. Suneson (9.8%) |
~59.5% | A lot of mutant alleles | Lee et al., 2021 | |||
|
GhFAD2-1A
GhFAD2-1D |
CRISPR/Cas9 | Ubiquitin (rice) | Agrobacterium-mediated transformation | No difference (Fibre quality/length/strength, micronaire, and germination) | 13.9% | 75.3–77.7% | m1-1 (−41 and +1 bp) m1-2 (+1 and −1 bp) m1-3 (+1 and +1 bp) m20-2 (−1 and +1 bp) m27-3 (−374 bp) |
Chen et al., 2021 |
| TaFAD2 | CRISPR/Cas9 | Ubiquitin4-2 (Petroselinum crispum) | Floral dipping | Late flowering, shorter plant height, low seed weight per plant, and low germination | 12% | ~35% |
fad2-4 (−2 bp) fad2-5 (+1 bp) fad2-6 (−29 bp) |
Jarvis et al., 2021 |
| OsFAD2-1 | CRISPR/Cas9 | Ubiquitin1 (maize) | Biolistic transformation | - |
Oryza sativa Japonica (No result) |
No result | +1 bp -302 bp |
Bahariah et al., 2021 |
| 2x 35S | Agrobacterium-mediated transformation | No difference |
Oryza sativa cv. Nipponbare (32%) |
~80% | 1–1 (+1 bp) 3–11 (+1 bp) 5–17 (−8 bp) 6–23 (−8 bp) |
Abe et al., 2018 | ||
| GmFAD2 | CRISPR/Cas9 | e35S | Agrobacterium-mediated transformation | - | cv. JN38 (17.34%) |
GmFAD2-2 (45.08–65.9%) | Substitution, −2, −3, +1, and +2 bp | Al Amin et al., 2019 |
| tipA | Agrobacterium-mediated transformation | No difference in plant height and grain weight. The grain is smaller and deeper in color |
cv. JN38 (19.15%) |
g3 strain (34.47%) g6 strain (40.45%) g36 strain (72.02%) |
GmFAD2-1A JN38g3–1 (+1 bp 66.7%) JN38g3–3 (−1 bp 16.6%) JN38g3–4 (−2 bp 16.7%) GmFAD2-2A JN38g6–2 (+1 bp 50%) JN38g6–3 (−1 bp 50%) Double JN38g36–3 (+1 and −1 bp 50%) JN38g36–5 (−2 and −7 bp 50%) |
Wu et al., 2020 | ||
| 2x 35S | Hairy root transformation | - | cv. Maverick (~20%) |
GmFAD2-1A, GmFAD2-1B homozygous lines (~80%) |
A lot of mutant alleles | Do et al., 2019 |