Table 1 ReMOT Control gene editing using optimized concentrations for EER and the RNPs Cas9, P2C-Cas9 and P2C-EGFP-Cas9s
Protein | Time of injection PBF | Individual injections (N) | Females laying KO | Females laying KO (%) | G0 larvae hatched (N) | G0 mosaic (N) | G0 white (N) | #KO/ injection EEF (%) | #KO/surviving G0 GEF (%) |
|---|---|---|---|---|---|---|---|---|---|
Maternal gene editing: wild-type females × white-eye males (mono-allelic) | |||||||||
Cas9 | 24 h | 83 | 1 | 1.20% | 1527 | 0 | 1 | 1.2% | 0.1% |
54 h | 55 | 0 | 0% | 1090 | 0 | 0 | 0.0% | 0.0% | |
Total | 138 | 1 | 0.70% | 2617 | 0 | 1 | 0.7% | 0.0% | |
P2C-Cas9 | 24 h | 88 | 2a | 2.20% | 1798 | 0 | 18 | 20.5% | 1.0% |
54 h | 54 | 9 | 16.60% | 1052 | 2 | 23 | 46.3% | 2.4% | |
Total | 142 | 11 | 8% | 2850 | 2 | 41 | 30.3% | 1.5% | |
P2C-EGFP-Cas9 | 24 h | 90 | 3 | 3.00% | 534 | 0 | 6 | 4.4% | 1.1% |
54 h | 134 | 5 | 2.90% | 1261 | 3 | 2 | 3.7% | 0.4% | |
Total | 224 | 8 | 4% | 1795 | 3 | 8 | 4.9% | 0.6% | |
Paternal gene editing: white eye (or heterozygous females) × wild-type males (mono and biallelic) | |||||||||
P2C-Cas9 | 54 h | 12 | 3 | 25% | 557 | 0 | 4 | 33.3% | 0.7% |
P2C-EGFP-Cas9 | 54 hb | 18 | 1 | 5.60% | 9 | 0 | 1 | 5.6% | 11.1% |
54 h | 40 | 2 | 5.00% | 213 | 2 | 0 | 5.0% | 0.9% | |
Total | 58 | 3 | 5.20% | 222 | 2 | 1 | 5.2% | 1.4% | |
Maternal and paternal gene editing: wild-type females × wild-type males (biallelic) | |||||||||
P2C-EGFP-Cas9 | 54 h | 58 | 1 | 1.70% | 1413 | 1 | 0 | 1.7% | 0.1% |
