Luciferase transfection in vivo

With the assistance of W. E. Balch (Scripps Research Institute, La Jolla, CA, USA), we created an adenoviral vector encoding destabilized luciferase (ad-Luc1) to allow transfection of lung tissue with luciferase adenoviral vector, which serves as a sensor of proteomic stress (21, 22). Mice were sedated with isoflurane and intubated using a 20-gauge angiocatheter. After confirmation of correct angiocatheter placement, ad-Luc1 virus (1 × 10¹⁰ plaque-forming units stock concentration), diluted in 50% dialysis buffer vehicle and 50% surfactant (Abbott Laboratories, Abbott Park, IL, USA) to achieve a dose of 1 × 10⁸ plaque-forming units in 50 µl/mouse, was intratracheally administered. To ensure survival of the animals, some were ventilated for ∼10 min after vector instillation (respiratory rate, 90 breaths/min; tidal volume, 150 μl; positive end–expiratory pressure, 5 cmH₂O; oxygen level, 100%).