Average vector copy number per transduced cell in the bone marrow of NOD/SCID mice as determined by semiquantitative PCR analysis
| Mouse no. . | hu genomes per 100 total genomes . | Vector copies per 100 genomes . | Vector copies per human cell . | TE FACS (%) . | Vector copy no. per GFP+ human cell . |
|---|---|---|---|---|---|
| PGK vector | |||||
| 1 | 18.6 | 6.17 | 0.33 | 16.0 | 2.1 |
| 2 | 13.5 | 1.70 | 0.13 | 3.0 | 4.2 |
| 3 | 17.8 | 1.37 | 0.08 | 1.6 | 4.8 |
| (selected mouse 7.23) | 7.1 | 46.1 | 6.47 | 91 | 6.5 |
| EF-1α SIN vector | |||||
| 11 | 8.5 | 1.09 | 0.13 | 4.0 | 3.2 |
| 12 | 11.3 | 2.82 | 0.25 | 7.7 | 3.3 |
| 13 | 20.0 | 6.01 | 0.30 | 4.9 | 6.1 |
| 14 | 26.0 | 4.32 | 0.17 | 5.3 | 3.1 |
| MESVltr vector | |||||
| 36 | 0.4 | 0.88 | 2.00 | 85 | 2.4 |
| 37 | 2.1 | 9.96 | 4.68 | 90 | 5.2 |
| 38 | 2.1 | 4.47 | 2.16 | 21 | 10.3 |
| CMV SIN vector | |||||
| 47 | 75.5 | 129.09 | 1.71 | 15.0 | 11.6 |
| 48 | 3.4 | 4.35 | 1.28 | 12.0 | 10.6 |
| Mouse no. . | hu genomes per 100 total genomes . | Vector copies per 100 genomes . | Vector copies per human cell . | TE FACS (%) . | Vector copy no. per GFP+ human cell . |
|---|---|---|---|---|---|
| PGK vector | |||||
| 1 | 18.6 | 6.17 | 0.33 | 16.0 | 2.1 |
| 2 | 13.5 | 1.70 | 0.13 | 3.0 | 4.2 |
| 3 | 17.8 | 1.37 | 0.08 | 1.6 | 4.8 |
| (selected mouse 7.23) | 7.1 | 46.1 | 6.47 | 91 | 6.5 |
| EF-1α SIN vector | |||||
| 11 | 8.5 | 1.09 | 0.13 | 4.0 | 3.2 |
| 12 | 11.3 | 2.82 | 0.25 | 7.7 | 3.3 |
| 13 | 20.0 | 6.01 | 0.30 | 4.9 | 6.1 |
| 14 | 26.0 | 4.32 | 0.17 | 5.3 | 3.1 |
| MESVltr vector | |||||
| 36 | 0.4 | 0.88 | 2.00 | 85 | 2.4 |
| 37 | 2.1 | 9.96 | 4.68 | 90 | 5.2 |
| 38 | 2.1 | 4.47 | 2.16 | 21 | 10.3 |
| CMV SIN vector | |||||
| 47 | 75.5 | 129.09 | 1.71 | 15.0 | 11.6 |
| 48 | 3.4 | 4.35 | 1.28 | 12.0 | 10.6 |
The results reveal multiple vector copies present per transduced cell, suggesting multiple lentiviral vector copy integration into NOD/SCID repopulating cells (from Figure 1). To calculate the copy number per transduced cell, the vector copy number per human cell was divided by the proportion of GFP+ cells as determined by FACS. The average vector copy number per transduced cell and the SD are shown for each vector group. TE indicates transduction efficiency. The average for the PGK vector (excluding selected mouse 7.23) was 3.7 ± 1.4; for the EF-1α SIN vector, 3.9 ± 1.5; for the MESVltr vector, 5.7 ± 4.0; for all groups, 5.6 ± 3.3.