Table 1.
Number of independent transgenic lines used for each species and genotyping of transgene-free T2 Plants.
| Species | Target | Number of Primary transformants | Number of independent lines (3:1 ratio) | Number of selected lines | Number of T2/T1 plants | Genotype T2/T1 plants | ||||
|---|---|---|---|---|---|---|---|---|---|---|
| Mut. Hom. | Biallelic | Heter. | wt | WT* | ||||||
| Oryza sativa | Os04g56950 | 20 | 14 | 14 | 31 | 20 (65%) | 8 (26%) | 2 ( 6%) | 1 (3%) | 1 (3%) |
| Solanum lycopersicum | Solyc07g64990 | 8 | 4 | 4 | 29 | 2 (7%) | 0 | 3 (10%) | 24 (83%) | 6 (21%) |
| Solyc12g14500 | 11 (38%) | 12 (41%) | 0 | 6 (21%) | ||||||
| Arabidopsis thaliana | At5g55250-1 | 15 | 12 | 4 | 214 | 0 | 0 | 3 (1.4%) | 211 (98.6%) | 112 (52%) |
| At5g55250-2 | 9 (4.2%) | 5 (2.3%) | 14 (6.5%) | 186 (86.9%) | ||||||
| At5g55250-3 | 42 (19.6%) | 2 (0.9%) | 43 (20.1%) | 127 (59.3%) | ||||||
Each genomic region was PCR amplified and sequencing to determine genotype of each genomic target and classify in four groups: both chromosomes mutated with the same mutation (Mut. Hom.); both chromosomes mutated but with different mutations (biallelic), only one chromosome with mutation (Heter.), and no detected mutation (wt). *The last column indicates de number of plants with wild-type sequence in all CRISPR targets.