%0 Journal Article
%T Production of lentiviral vectors using novel, enzymatically produced, linear DNA
%J -
%D 2019
%R https://doi.org/10.1038/s41434-018-0056-1
%X The manufacture of large quantities of high-quality DNA is a major bottleneck in the production of viral vectors for gene therapy. Touchlight Genetics has developed a proprietary abiological technology that addresses the major issues in commercial DNA supply. The technology uses ¡®rolling-circle¡¯ amplification to produce large quantities of concatameric DNA that is then processed to create closed linear double-stranded DNA by enzymatic digestion. This novel form of DNA, Doggybone£¿ DNA (dbDNA£¿), is structurally distinct from plasmid DNA. Here we compare lentiviral vectors production from dbDNA£¿ and plasmid DNA. Lentiviral vectors were administered to neonatal mice via intracerebroventricular injection. Luciferase expression was quantified in conscious mice continually by whole-body bioluminescent imaging. We observed long-term luciferase expression using dbDNA£¿-derived vectors, which was comparable to plasmid-derived lentivirus vectors. Here we have demonstrated that functional lentiviral vectors can be produced using the novel dbDNA£¿ configuration for delivery in vitro and in vivo. Importantly, this could enable lentiviral vector packaging of complex DNA sequences that have previously been incompatible with bacterial propagation systems, as dbDNA£¿ technology could circumvent such restrictions through its phi29-based rolling-circle amplification
%U https://www.nature.com/articles/s41434-018-0056-1