Table 1.
Optimization of cpRPE65-mCherry01 F0 generation transgenic newts
| DNA pga/egg | Eggs injectedb | Survival St.10 blastula (%) | mCherry expression | Survivald St. 59 (%) | ||||
|---|---|---|---|---|---|---|---|---|
| Developing eye (%) | St. 59 RPEc | |||||||
| ND | Avg. | Strong | + | – | ||||
| 160 | 120 | 22 (18) | 8 | 11 (50) | 3 (13.6) | 4/5 | 1/5 | 9 (41) |
| 100 | 112 | 45 (40) | 32 | 9 (20) | 4 (9) | 3/3 | 0/3 | 23 (47) |
| 80 | 103 | 68 (66) | 60 | 6 (9) | 2 (3) | 2/2 | 0/2 | 46 (68) |
| 40 | 108 | 77 (71) | 74 | 3 (4) | 0 | 2/2 | 0/2 | 64 (83) |
| 0 | – | 64e (97) | – | – | – | – | – | 59 (92) |
ND not detected, St. stage, RPE retinal pigment epithelium, Avg. average
apg is the total mass of cpRPE65-mCherry01 including the vector backbone
bThe injection volume was fixed at 2 nL/egg. I-SceI enzyme concentration was fixed at 1× 10–3 U/egg
cA subset of the average mCherry-expressing embryos were reared until stage 59 and their eye sections were prepared as shown in Fig. 3j–o
dBased on surviving blastula embryos
eViability control group (n = 66)