Table 1.
T0 Fielder wheat plants | |||
---|---|---|---|
Regenerated | Expressing Cas9 | 1 or 2 transgene copies | |
Construct α1_sgRNA sgRNA_α87 |
40 | 38 (95%) | 21 (55%) |
Construct α2_sgRNA sgRNA_α213 + sgRNA_α324 |
32 | 12 (38%) | 4 (33%) |
Construct γ3_sgRNA sgRNA_γ86 + sgRNA_γ272 + sgRNA_γ603 |
36 | 35 (97%) | 26 (74%) |
Construct α2γ3_sgRNA sgRNA_α213 + sgRNA_α324 + sgRNA_γ86 + sgRNA_γ272 + sgRNA_γ603 |
42 | 32* (76%) | 12 (38%) |
TOTAL | 150 | 117 (78%) | 63 (54%) |
For each construct, between 32 and 42 plants were regenerated. The * indicates that 34 plants expressed Cas9 but 2 were discarded since they did not contain all 5 sgRNAs. Overall, 78% of plants expressed the Cas9, among which 54% had 1 or 2 copies of the nptII selectable marker gene whereas the remainder contained additional copies of nptII.