Minimal DNA vectors exclusively comprising therapeutically relevant sequences hold great promise for the development of novel therapeutic regimen.Dumbbell-shaped vectors represent non-viral non-integrating DNA minimal vectors which have entered an advanced stage of clinical development (Hardee et al., 2017).
Spliceable introns and DNA Miele DA3496 89 cm Canopy Cooker Hood - Stainless Steel nuclear import signals such as SV40 enhancer sequences are molecular features that have found multiple applications in plasmid vectors to improve transgene expression.In dumbbells however, effects triggered by introns were not investigated and DNA-based nuclear import sequences have not found applications yet, presumably because dumbbell vectors have continuously been minimized with regard to size.We investigated the effects of an intron and/or SV40 enhancer derived sequences on dumbbell vector driven reporter gene expression.
The implementation of a spliceable intron was found to enhance gene expression unconditionally in all investigated cell lines.Conversely, the use of the SV40 enhancer improved gene expression in a cell type-dependent manner.Though both features significantly enlarge dumbbell vector size, neither the intron nor the enhancer or a combination of both revealed a negative effect on gene expression.
On the contrary, both features together improved dumbbell-driven gene expression up to 160- or 56-fold compared with plasmids or control dumbbells.Thus, it is LAVENDER BATH highly recommended to consider an intron and the SV40 enhancer for dumbbell vector design.Such an advanced design can facilitate pre-clinical and clinical applications of dumbbell-shaped DNA vectors.