Table 3.
PCE correlates with reduced cytoplasmic accumulation of luc RNA in transduced cells.
| RNA copy number (× 103)a | ||||||
| Nucleoplasm | Cytoplasm | Cytoplasmic accumulationb | Translational efficiencyc | |||
| Vector | luc | actin | luc | actin | ||
| U3-Luc | 56.7 ± 5.4 (1) | 59.3 | 25.3 ± 2.8 (1) | 375 | 0.45 | 1 |
| PCE-Luc | 34.9 ± 3.8 (0.47) | 76.6 | 5.4 ± 0.2 (0.2) | 372 | 0.16 | 5 |
| IE-U3-Luc | 91.3 ± 1.5 (1) | 69.1 | 46.8 ± 12.9 (1) | 417 | 0.51 | 0.5 |
| IE-PCE-Luc | 96.8 ± 2.7 (0.88) | 82.5 | 18.3 ± 2.7 (0.4) | 384 | 0.19 | 3 |
a Equivalent amounts (100 ng) of RNA from either the nucleoplasm or cytoplasm were reverse transcribed to generate cDNA and one-tenth of each reaction was quantified by real-time PCR with primers specific to luc or actin. Copy numbers were derived from standard curves generated with pGL3 luciferase plasmid in the range of 101 to 109 copies. Reactions were performed in duplicate and the mean and standard deviation are indicated. (), levels of luc normalized to actin relative to PCE-lacking controls, U3-Luc or IE-U3-Luc, respectively.
b Ratio of the copy number of luc RNA in the cytoplasm and nucleoplasm.
c Ratio of Luc activity to copy number of cytoplasmic luc RNA