Table 2.
Bimolecular integration assay in human cells
| Plasmid | Blues/total | % Integration | Corrected % integration |
|---|---|---|---|
| p220Kan | |||
| No DNA | 0/7,970 | <0.01 | <0.08 |
| pCMVInt only | 0/13,600 | <0.007 | <0.04 |
| pTSAD only | 1/11,830 | 0.008 | 0.05 |
| pCMVInt + pTSAD | 3/32,680 | 0.009 | 0.06 |
| p220KattB35 | |||
| No DNA | 0/64,530 | <0.002 | <0.01 |
| pCMVInt only | 0/42,720 | <0.002 | <0.02 |
| pTSAD only | 0/39,930 | <0.003 | <0.02 |
| pCMVInt + pTSAD | 382/157,710 | 0.242 | 1.69 |
| p220KattBfull | |||
| No DNA | 0/70,350 | <0.001 | <0.01 |
| pCMVInt only | 0/41,960 | <0.002 | <0.02 |
| pTSAD only | 0/39,740 | <0.003 | <0.02 |
| pCMVInt + pTSAD | 1,799/166,890 | 1.08 | 7.50 |
Results are shown for human cells carrying one of three EBV plasmids, p220Kan, a negative control lacking attB; p220KattB35, which carries a similar attB; and p220KattBfull, carrying the full-sized attB. Integration frequencies are shown for experiments when no DNA was transfected, when either the integrase expression plasmid pCMVInt or the attP-bearing plasmid pTSAD were transfected alone, or when both pCMVInt and pTSAD were transfected together. Only the latter condition, in the presence of a plasmid bearing attB, led to integration events. Integration frequencies were corrected for transfection frequency to give the accurate corrected integration frequencies in the last column. p220KattBfull produced the highest integration frequency at 7.5%.