Overview of experiments performed in this study
Vector, MOI . | Titer of supernatant, 106/mL . | Treated cells . | Positive wells on 96-well plate . | Frequency of replating cells . | No. clones of analyzed samples . | Mean copy no., d 7 qPCR . |
|---|---|---|---|---|---|---|
| Pilot experiment | ||||||
| LTRSFdsRED, 5 | 3 | 1 × 106 | 56; 81 | 1 in 80 | 1 ; 3 | ND |
| Experiment 1 | ||||||
| LTRSFeGFP, 20 | 15 | 1 × 105 | 50; 70 | 1 in 102 | 2 ; 9 | ND |
| SINSFeGFP, 20 | 5 | 1 × 105 | 2; 3 | 1 in 3790 | ND | ND |
| Mock, 0 | — | 1 × 105 | 4; 5 | < 1 in 2083 | ND | ND |
| Experiment 2 | ||||||
| LTRSFeGFP, 20 | 20 | 1 × 105 | 96; 96 | > 1 in 22 | 2 ; 4 | 11.6 |
| SINSFeGFP, 20 | 5 | 1 × 105 | 0; 6 | 1 in 3150 | ND | 2.9 |
| LTRSFdsRed, 20 | 23 | 1 × 105 | 96; 96 | > 1 in 22 | 2 ; 4 | 13.9 |
| SINSFdsRed, 20 | 5 | 1 × 105 | 0; 0 | < 1 in 9550 | ND | 5.4 |
| Mock, 0 | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
| Experiment 3 | ||||||
| LTRSFeGFP, 20 | 8 | 1 × 105 | 52; 40 | 1 in 153 | 1 ; 10 | 3 |
| SINSFeGFP, 20 | 9 | 1 × 105 | 1; 0 | < 1 in 9550 | ND | 3.1 |
| Mock, 0 | — | 1 × 105 | 0; 0 | < 1 in 9550 | ND | ND |
| Experiment 4 | ||||||
| LTRSFeGFP, 20 | 9 | 1 × 105 | 42; 40 | 1 in 180 | 3 ; 3 | 6.4 |
| LTRSFeGFP, 10 | 9 | 1 × 105 | 20 | 1 in 428 | ND | 4.2 |
| LTRSFeGFP, 5 | 9 | 1 × 105 | 15 | 1 in 589 | ND | 2.2 |
| LTRSFeGFP, 2.5 | 9 | 1 × 105 | 15 | 1 in 589 | ND | 0.9 |
| SINSFeGFP, 20 | 10 | 1 × 105 | 32; 34 | 1 in 237 | 2 ; 6 | 4.4 |
| Mock, 0 | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
| Experiment 5 | ||||||
| LTRSFeGFP, 20 | 10 | 1 × 105 | 96 | > 1 in 22 | ND | 4 |
| LTRSFeGFP, 5 | 10 | 1 × 105 | 31 | 1 in 256 | ND | 1.5 |
| LTRSFeGFP, 1.25 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.6 |
| SINSFeGFP, 20 | 10 | 1 × 105 | 13 | 1 in 687 | 1 ; 5 | 1.7 |
| SINSFeGFP, 5 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.6 |
| SINSFeGFP, 1.25 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.5 |
| Mock | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
Vector, MOI . | Titer of supernatant, 106/mL . | Treated cells . | Positive wells on 96-well plate . | Frequency of replating cells . | No. clones of analyzed samples . | Mean copy no., d 7 qPCR . |
|---|---|---|---|---|---|---|
| Pilot experiment | ||||||
| LTRSFdsRED, 5 | 3 | 1 × 106 | 56; 81 | 1 in 80 | 1 ; 3 | ND |
| Experiment 1 | ||||||
| LTRSFeGFP, 20 | 15 | 1 × 105 | 50; 70 | 1 in 102 | 2 ; 9 | ND |
| SINSFeGFP, 20 | 5 | 1 × 105 | 2; 3 | 1 in 3790 | ND | ND |
| Mock, 0 | — | 1 × 105 | 4; 5 | < 1 in 2083 | ND | ND |
| Experiment 2 | ||||||
| LTRSFeGFP, 20 | 20 | 1 × 105 | 96; 96 | > 1 in 22 | 2 ; 4 | 11.6 |
| SINSFeGFP, 20 | 5 | 1 × 105 | 0; 6 | 1 in 3150 | ND | 2.9 |
| LTRSFdsRed, 20 | 23 | 1 × 105 | 96; 96 | > 1 in 22 | 2 ; 4 | 13.9 |
| SINSFdsRed, 20 | 5 | 1 × 105 | 0; 0 | < 1 in 9550 | ND | 5.4 |
| Mock, 0 | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
| Experiment 3 | ||||||
| LTRSFeGFP, 20 | 8 | 1 × 105 | 52; 40 | 1 in 153 | 1 ; 10 | 3 |
| SINSFeGFP, 20 | 9 | 1 × 105 | 1; 0 | < 1 in 9550 | ND | 3.1 |
| Mock, 0 | — | 1 × 105 | 0; 0 | < 1 in 9550 | ND | ND |
| Experiment 4 | ||||||
| LTRSFeGFP, 20 | 9 | 1 × 105 | 42; 40 | 1 in 180 | 3 ; 3 | 6.4 |
| LTRSFeGFP, 10 | 9 | 1 × 105 | 20 | 1 in 428 | ND | 4.2 |
| LTRSFeGFP, 5 | 9 | 1 × 105 | 15 | 1 in 589 | ND | 2.2 |
| LTRSFeGFP, 2.5 | 9 | 1 × 105 | 15 | 1 in 589 | ND | 0.9 |
| SINSFeGFP, 20 | 10 | 1 × 105 | 32; 34 | 1 in 237 | 2 ; 6 | 4.4 |
| Mock, 0 | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
| Experiment 5 | ||||||
| LTRSFeGFP, 20 | 10 | 1 × 105 | 96 | > 1 in 22 | ND | 4 |
| LTRSFeGFP, 5 | 10 | 1 × 105 | 31 | 1 in 256 | ND | 1.5 |
| LTRSFeGFP, 1.25 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.6 |
| SINSFeGFP, 20 | 10 | 1 × 105 | 13 | 1 in 687 | 1 ; 5 | 1.7 |
| SINSFeGFP, 5 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.6 |
| SINSFeGFP, 1.25 | 10 | 1 × 105 | 0 | < 1 in 9550 | ND | 0.5 |
| Mock | — | 1 × 105 | 0 | < 1 in 9550 | ND | ND |
Titer was determined by limiting dilution on SC-1 fibroblasts and was adjusted before the transduction ; bone marrow cells. Pairs of numbers separated by semicolons result from duplicate determinations
ND indicates not determined; —, not applicable