Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells
Marie Soukupovà, Silvia Zucchini, Pascal Trempat, Selene Ingusci, Coline Perrier-Biollay, Mario Barbieri, Stefano Cattaneo, Barbara Bettegazzi, Simonetta Falzoni, Hervé Berthommé, Michele Simonato
Abstract
using different promoters and preventing transgene silencing. To pursue the latter, we deleted bacterial DNA sequences derived from vector construction and shielded the transgene cassette using AT-rich and insulator-like sequences (SAm technology). We employed luciferase and GFP as reporter genes. To determine transgene expression kinetics, we injected vectors in the hippocampus of mice that were longitudinally scanned for bioluminescence for 6 months. To evaluate safety, we analyzed multiple markers of damage and performed patch clamp electrophysiology experiments. All vectors proved safe, and we managed to modulate the duration of transgene expression, up to obtaining a stable, long-lasting expression using the SAm technology. Therefore, these amplicon vectors represent a flexible, efficient, and safe tool for gene delivery in the brain.