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Performance of Ad-A-Gene EGFP-Glucocorticoid Receptor (EGFP-GCCR): an adenoviral vector gene delivery system

e response (EC50 values) to dexamethasone in transduced cells compared favourably to that in stable cell lines, and similar Hill slope values were generated that indicate single-binding site stoichiometry in both cases.

Note: Setup of this experiment required use of different cell types and this should be considered when attempting to make direct comparisons of the resulting data.

Reproducibility of adenoviral transduction
Figure 7 shows normalized results obtained from three experiments in which transduced HeLa cells transiently expressing EGFP-GCCR were treated with varying doses of dexamethasone. The data show that adenoviral transduction is both robust and reproducible and that there is excellent agreement in EC50 values across the three experiments. Goodness-of-fit (R2) is well within acceptable limits.


Conclusion
The EGFP-GCCR chimera, when transiently expressed in host cells after adenoviral transduction, functions well in a translocation assay upon exposure to pharmacologically relevant stimuli. The response is both time and dose dependent. Use of the EGFP-GCCR adenoviral vector shows broad tropism of transduction in a range of target cells with subsequent assay results comparing favorably to those obtained in a stable cell line expressing GFP-tagged GCCR. Experimental variation, when carrying out adenoviral transduction followed by functional evaluation, is statistically acceptable.

The sequential process of viral transduction, exposure of cells to reagents, and data acquisition can be transcribed into suitable automated and integrated workflow systems with standard liquid-handling dispensing units as well as image capture and data analysis workstations.


References
1. Zhang, S. and Daniellsen, M. Steroid structures and activities [Online] http://nrr.georgetown.edu/NRR
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