Table 2 Relative cleavage activity of RGEN and TALEN constructs in transiently transformed barley and tobacco leaf explants
From: A simple test for the cleavage activity of customized endonucleases in plants
Plant species | Target gene | Type of customized endonuclease | Constructs used | Experiment | YFP cells | mCherry cells | Ratio YFP/mCherry cells (%) |
|---|---|---|---|---|---|---|---|
Tobacco | gfp | RGEN | pTARGET-gfp1 | 1 | 273 | 371 | 73.6 |
+ RGEN-gfp | 2 | 342 | 389 | 87.9 | |||
| Â | 3 | 324 | 499 | 64.9 | |||
|  | Average | 313 ± 29 | 420 ± 57 | 74.6 | |||
pTARGET-gfp1 | 1 | 0 | 106 | 0 | |||
| Â | 2 | 0 | 204 | 0 | |||
| Â | 3 | 0 | 81 | 0 | |||
|  | Average | 0 | 130 ± 65 | 0 | |||
TALEN | pTARGET-gfp1 | 1 | 195 | 361 | 54.0 | ||
+ TALEN-gfp | 2 | 25 | 183 | 13.7 | |||
| Â | 3 | 80 | 545 | 14.7 | |||
|  | Average | 100 ± 71 | 363 ± 148 | 27.5 | |||
Barley | gfp | RGEN | pTARGET-gfp2 | 1 | 72 | 207 | 34.8 |
+ RGEN-gfp | 2 | 55 | 206 | 26.7 | |||
| Â | 3 | 83 | 255 | 32.5 | |||
|  | Average | 70 ± 12 | 223 ± 23 | 31.4 | |||
pTARGET-gfp2 | 1 | 0 | 46 | 0 | |||
TALEN | pTARGET-gfp1 | 1 | 71 | 223 | 31.8 | ||
+ TALEN-gfp | 2 | 110 | 331 | 33.2 | |||
| Â | 3 | 49 | 195 | 25.1 | |||
|  | Average | 77 ± 25 | 250 ± 59 | 30.7 | |||
pTARGET-gfp1 | 1 | 0 | 44 | 0 | |||
MLO | TALEN | pTARGET-MLO | 1 | 134 | 350 | 38.3 | |
+ TALEN-MLO | 2 | 107 | 254 | 42.1 | |||
| Â | 3 | 112 | 237 | 47.3 | |||
|  | Average | 118 ± 12 | 280 ± 50 | 42.0 | |||
pTARGET-MLO | 1 | 0 | 52 | 0 |