neDNA™ Is A Robust Alternative To Plasmid DNA For AAV Production

By Navarro A, Maestro S, Etxabe A, Arranz N, Isábal M, Alcalá M, Silva E, Mora L, Cristóbal A, Iribar H, Lang V, Guarinoni T, Moullier P, Trigueros C, and François A

While the transfection of three plasmids – one carrying the desired gene flanked by AAV inverted terminal repeats, another encoding the AAV rep and cap genes, and a third encoding adenovirus helper functions – remains the most common method for producing recombinant adeno-associated virus (AAV) vectors, this approach suffers from a drawback: plasmid DNA can contaminate the final AAV product with bacterial sequences, potentially raising safety concerns. This study investigates neDNA™, a promising alternative. NeDNA™ consists of linear DNA molecules with sealed ends, devoid of bacterial sequences. We directly compared plasmids and neDNA™ for AAV production in Pro10™ cells, employing various AAV capsid serotypes and production scales. Additionally, we assessed the potency of the resulting vectors in an in vivo study conducted in mice.