Table 3.
CRISPR/Zygote Approach Microinjection Results.
| Experiment Number | Donor Vector (ng/µL) | Guide Set | Zygotes Injected/Transferred | Newborns | Liveborna | Stillbornb | Lost Before Weanc | Weanedd | Founderse | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 and 6 combined | 5 | OPTIMAL POSITION (G5-G8) | 147 | 33 | 22.4% | 30 | 90.9% | 3 | 9.1% | 1 | 3.3%% | 29 | 96.7% | 1f | 3.4% |
| 2 | 1 | OPTIMAL POSITION (G5-G8) | 75 | 8 | 10.7% | 8 | 100.0% | 0 | 0.0% | 0 | 0.0% | 8 | 100.0% | 1g | 12.5% |
| 4 | 1 | OPTIMAL SCORE (G1-G4) | 81 | 17 | 21.0% | 17 | 100.0% | 0 | 0.0% | 1 | 5.9% | 16 | 94.1% | 0 | 0.0% |
| 7 | 10 | OPTIMAL SCORE (G1-G4) | 69 | 8 | 11.6% | 7 | 87.5% | 1 | 12.5% | 0 | 0.0% | 7 | 100.0% | 1g | 14.3% |
| 8 | 5 | OPTIMAL SCORE (G1-G4) | 76 | 23 | 30.3% | 22 | 95.7% | 1 | 4.3% | 1 | 4.5% | 21 | 95.5% | 0 | 0.0% |
| Totals | 448 | 89 | 19.9% | 84 | 94.4% | 5 | 5.6% | 3 | 3.6% | 81 | 96.4% | 3 | 3.7% | ||
Mice recovered from the CRISPR/Zygote Approach microinjection experiments (described in Table 2) are shown. These experiments tested permutations of two design parameters — targeting vector/donor DNA concentration (at 1, or 5, or 10 ng/µL) and sgRNA design (optimal score or optimal position). Experiments 1 and 6 are replicates; the results of these experiments are combined. Experiments 3 and 5 were technical failures resulting in no wean-age mice (see Supplementary Table 4 for details). Experiments 2, 6, and 7 generated single founder animals. See Table 4 for details.
aPercentage calculated as Liveborn/Newborns × 100.
bPercentage calculated as Stillborn/Newborns × 100.
cPercentage calculated as Lost Before Wean/Liveborn × 100.
dPercentage calculated as Weaned/Liveborn × 100.
ePercentage calculated as Founders/Weaned × 100.
fProper integration and transmission confirmed. See Tables 4, 5, and text for details.
gWas not transmitted through the germline. See Tables 4, 5, and text for details.