CFC assays of transduced CD34+ cells
Experiment . | Vector-positive CFCs, % . | Puromycin-resistant CFCs, % . | Trapping efficiency, % . |
|---|---|---|---|
| LT exp 1 | 23 | 0.5 | 2 |
| RT exp 1 | 18 | 3.3 | 18 |
| LT exp 2 | 96 | 7.1 | < 7* |
| RT exp 2 | 23 | 4.9 | 21 |
Experiment . | Vector-positive CFCs, % . | Puromycin-resistant CFCs, % . | Trapping efficiency, % . |
|---|---|---|---|
| LT exp 1 | 23 | 0.5 | 2 |
| RT exp 1 | 18 | 3.3 | 18 |
| LT exp 2 | 96 | 7.1 | < 7* |
| RT exp 2 | 23 | 4.9 | 21 |
Colony-forming cell (CFC) assays of the same cells shown in Figure 4B (exp1) and from a different experiment that used a higher input of LT vector (exp2), plated in semisolid medium with or without puromycin. The frequency of vector- positive CFCs was determined by PCR performed on unselected CFCs. The frequency of puromycin-resistant CFCs was calculated from the ratio between CFCs grown in medium with and without puromycin. The frequency of promoter trapping was calculated as the ratio between puromycin-resistant CFCs and vector-positive CFCs.
In the presence of multicopy vector integration, the frequency of trapping (trapping efficiency) is overestimated