Analyses performed on the unsorted bulk-transduced population
| Vector used to transduce K562-X-CGD cells . | Insert copy no. per gp91phox- positive K562-X-CGD cell . | Vector mRNA transcripts per genomic insert copy . | Mean fluorescence intensity of the gp91phox-positive cell population by flow cytometry . | PMA stimulated oxidase activity (LU/40 min/105gp91phox-positive cells) . |
|---|---|---|---|---|
| MFGS-gp91phox | 3.7 | 1427 | 383 | 1988 |
| Lenti-CMV-gp91phox | 3.2 | 176 | 58 | 508 |
| Lenti-EF1α-gp91phox | 4.4 | 1328 | 214 | 1815 |
| Vector used to transduce K562-X-CGD cells . | Insert copy no. per gp91phox- positive K562-X-CGD cell . | Vector mRNA transcripts per genomic insert copy . | Mean fluorescence intensity of the gp91phox-positive cell population by flow cytometry . | PMA stimulated oxidase activity (LU/40 min/105gp91phox-positive cells) . |
|---|---|---|---|---|
| MFGS-gp91phox | 3.7 | 1427 | 383 | 1988 |
| Lenti-CMV-gp91phox | 3.2 | 176 | 58 | 508 |
| Lenti-EF1α-gp91phox | 4.4 | 1328 | 214 | 1815 |
More than 70% of cells in all 3 transduction groups expressed gp91phox transgene. Identical results were found when gp91phox-expressing cells were purified by flow cytometry sorting first, and then insert copy number analyses and other determinations were performed. Not shown is that there was no improvement of expression levels when PGK replaced CMV promoter in the lentivector–gp91phox.