Digital imaging microscopy of firefly luciferase activity to directly monitor differences in cell transduction efficiencies between AdCMVLuc and Ad5LucRGD vectors having different cell binding properties

The luciferase reporter gene incorporated into adenoviral vectors is very u seful for monitoring viral transduction of different cell types or for comp aring the transduction efficiency of different viral constructs of one cell type. Luciferase protein expression can be detected and quantified with ve ry high sensitivity from whole cells or organ extracts. However. its disadv antages become obvious when aiming at evaluation of transduction events at the single cell level. The results obtained from whole cell extracts cannot be directly correlated to single cell events. In this paper direct cellula r luciferase imaging using cell permeable luciferin substrates is applied f or comparative analysis of cellular transduction events by two adenoviral v ectors with different cell binding properties. Using digital imaging micros copy we show a more than ten-fold increase in transduction efficiency by Ad 5LucRGD vectors versus AdCMVLuc vectors on human A549 cells. (C) 2001 Elsev ier Science B.V. All rights reserved.