Reduction of plasmid vector backbone length enhances reporter gene expression

Influence of DNA characteristics on cell membrane damage stimulated by electrical short-circuiting via a low-conductive aqueous droplet in dielectric oil

“…They reported that DNA aggregate formation was observed not only for large DNA molecules of 1000 bp but also for short DNA molecules less than 100 bp. Recently, Boye et al reported that reducing plasmid DNA backbone length significantly increased transgene expression levels, and this effect was observed in both electroporation and a commercially available transfection reagent [33]. Although further investigations are required to prove the detailed mechanism observed here, this study may have implications for understanding the transportation mechanism of electroporation.…”

The development of DNA vaccines against SARS-CoV-2

“…While it is true that the introduction of introns was reported to have a positive impact on plasmid optimization in terms of increased gene expression, there is also evidence that a larger size of the insert in the DNA plasmid vector might have a negative impact on the levels of gene expression. In an experiment where two vectors of differing plasmid lengths containing a firefly luciferase encoding gene insert were compared for levels of gene expression, it was observed that 2-fold decrease in the size of the plasmid vector backbone could increase gene expression levels by more than 10-fold in rat tenocytes in vitro, and rat myocardium in vivo [ 83 ] . Moreover, the administration of varying sizes of plasmid DNA into Bacillus subtilis ISW1214 showed that transformation efficiency was reduced with increasing size of the plasmid DNA [ 84 ].…”

Improving cell and gene therapy safety and performance using next-generation Nanoplasmid vectors

“…Robust transgene expression up to 10-fold higher than plasmid vectors has been reported with Nanoplasmid vectors after in vitro delivery to rat tendon or mouse melanoma cells, or in vivo electroporation delivery to rat skin or heart ( Figure 3 ). 61 , 62 Nanoplasmid has also shown robust expression after nanoparticle tail vein delivery, 63 hydrodynamic retrograde intrabiliary injection in pigs, 64 and after nanoparticle delivery to tumor cells in vitro and in vivo, 65 or electroporation delivery of DNA-encoded antibody therapeutics to muscle. 66 An immune-oncology gene therapy named EG-70 (also known as detalimogen voraplasmid) is currently in a phase I/II trial for BCG refractory non-muscle invasive bladder cancer (NCT04752722).…”

“…(∗=p<0.01, ∗∗= p<0.001) This figure was adapted from Boye et al. 61 “Reduction of plasmid vector backbone length enhances reporter gene expression,” 2022, Bioelectrochemistry, 144, 107981, https://doi.org/10.1016/j.bioelechem.2021.107981 CC BY-NC -ND 4.0. …”