Investigations of the effect of DNA size in transient transfection assay using dual luciferase system

Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy

“…MNPs have emerged strongly as a class of advanced functional materials for neuro-regeneration, serving as 'multifunctional tools' for cell therapy given additional applications for non-invasive cell imaging and magnetic cell targeting, so it is clear that this non-viral gene transfer method offers significant benefits for clinical translation [26]. Despite the critical advantages offered by this technique, we and others have reported that DNA plasmid size bears an inverse relationship with transfection efficiency for non-viral methods such as liposome-and nanoparticle-mediated transfection [23,[27][28][29]. For example, using lipofection, a systematic analysis of luciferase expression levels (using reporter constructs of increasing size; 4.8 kb to 10.5 kb) showed reduced transfection with greater plasmid size [29].…”

Nanoscale structure of protamine/DNA complexes for gene delivery

“…The existence of an inverse correlation between DNA size and transfection efficiency has been corroborated by recent results. [15][16][17] Still, a well-assessed interpretation is lacking. Whatever the involved mechanism, it is now well established that all transfection barriers are strongly affected by the ultrastructural features of DNAbased complexes.…”

Preferential Epigenetic Suppression of the Autonomous MusD over the Nonautonomous ETn Mouse Retrotransposons

“…With large DNA fragments introduced into plasmid DNA, we saw evidence of reduced plasmid DNA uptake via lipofection. Such effects have been described previously and shown to account for greater-than-10-fold reductions in luciferase expression, inversely correlating with insert size (69). We cloned a 3-kb and a 3.7-kb lambda phage "stuffer" DNA fragment into the SalI restriction site of the pGL3B vector to serve as a promoterless size-adjusted control and normalized pGL3B-LTR#7 luciferase activity to nonmodified pGL3B, since the LTR is only 317 bp long, while the luciferase activities of the 3.15-kb ETnII internal region-containing constructs were (38).…”