Table 2.
Mutation rates in Zea mays IPK gene target region produced by CRISPR-Cas RNP delivery and analyzed by Sanger sequencing and ICE software analysis.
| Sample | Incubation Time (min) | % of Indel | Model Fit (R2) | KO Score | Mutation Range (bp) | Greater Contribution (bp) |
|---|---|---|---|---|---|---|
| Cas9 only | 20 | 0 | 0 | 0 | 0 | |
| gRNA2 only | 20 | 0 | 0 | 0 | 0 | |
| gRNA3 only | 20 | 0 | 0 | 0 | 0 | |
| gRNA4 only | 20 | 0 | 0 | 0 | 0 | |
| Cas9 + gRNA2 rep1 | 20 | 0 | 0.99 | 0 | 0 | 0 |
| Cas9 + gRNA2 rep2 | 20 | 1 | 1 | 1 | −4 to −2 | −2 |
| Cas9 + gRNA3 rep1 | 20 | 0 | 1 | 0 | −7 to −1 | −2 |
| Cas9 + gRNA3 rep2 | 20 | 1 | 1 | 1 | −7 to−1 | −2 |
| Cas9 + gRNA4 rep1 | 20 | 3 | 0.99 | 2 | −7 to +12 | −2 |
| Cas9 + gRNA4 rep2 | 20 | 1 | 0.99 | 1 | −2 to +3 | −2 |
| Cas9 + gRNA2 rep1 | 40 | 4 | 0.99 | 4 | −7 to +1 | −1 |
| Cas9 + gRNA2 rep2 | 40 | 6 | 1 | 6 | -8 to +1 | +1 |
| Cas9 + gRNA3 rep1 | 40 | 1 | 1 | 1 | −19 to−1 | −1 |
| Cas9 + gRNA3 rep2 | 40 | 2 | 0.96 | 2 | −28 to +8 | −19 |
| Cas9 + gRNA4 rep1 | 40 | 2 | 0.99 | 2 | −3 to −2 | −2 |
| Cas9 + gRNA4 rep2 | 40 | 5 | 1 | 4 | −16 to +12 | −2 |
Note: KO = Knockout.