Table 1. Generation of Il11 mutant mice using vitrified/warmed zygotes.
| Reagent | Pulse | Age in weeks | Culture time | Electroporated | Recovered | Transferred(2-cell embryo) | Pups (%) | Transferred(1-cell zygote) | Pups (%) | HMA+* | Sequencing+** | Single-peak mutation*** | Mutants (%) |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| IL11_B RNP (500 ng/µl Cas9 and 250 ng/µl gRNA) | 25 V × 7 | 4 | 1 h | 52 | 51 | 46 | 7 (15.2) | – | – | 6 | 1 | 0 | 7 (100) |
| 2 h | 27 | 27 | 25 | 6 (24.0) | – | – | 4 | 2 | 0 | 6 (100) | |||
| 3 h | 24 | 24 | 22 | 5 (22.7) | – | – | 3 | 2 | 2 | 5 (100) | |||
| 5 h | 50 | 50 | 47 | 16 (34.0) | – | – | 14 | 2 | 1 | 16 (100) | |||
| 2 h | 46 | 46 | – | – | 46 | 7 (15.2) | 6 | 1 | 1 | 7 (100) | |||
| 5 h | 31 | 31 | – | – | 31 | 2 (6.5) | 1 | 0 | 0 | 1 (50.0) | |||
| 11–12 | 1 h | 29 | 29 | 28 | 4 (14.3) | – | – | 3 | 1 | 0 | 4 (100) | ||
| 3 h | 24 | 24 | 24 | 8 (33.3) | – | – | 7 | 1 | 0 | 8 (100) | |||
| 5 h | 23 | 23 | 20 | 7 (35.0) | – | – | 7 | – | – | 7 (100) | |||
| 2 h | 63 | 61 | – | – | 61 | 17 (27.9) | 11 | 5 | 1 | 16 (94.1) | |||
| 5 h | 29 | 29 | – | – | 29 | 5 (17.2) | 4 | 1 | 0 | 5 (100) | |||
*The numbers of HMA-positive founders are shown. **The numbers of mutant mice identified by sequencing analysis are shown. ***The numbers of pups containing single-peak mutation analyzed by direct sequencing analysis are shown.