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Towards effective non-viral gene delivery vector

Šimčíková, Michaela, Prather, Kristala L. J., Prazeres, Duarte M. F., Monteiro, Gabriel A.
Biotechnology & genetic engineering reviews 2015 v.31 no.1-2 pp. 82-107
bioavailability, biopharmaceuticals, biotechnology, clinical trials, gene expression, genetic engineering, introns, manufacturing, plasmids, protein synthesis, replication origin, therapeutics, transfection
Despite very good safety records, clinical trials using plasmid DNA failed due to low transfection efficiency and brief transgene expression. Although this failure is both due to poor plasmid design and to inefficient delivery methods, here we will focus on the former. The DNA elements like CpG motifs, selection markers, origins of replication, cryptic eukaryotic signals or nuclease-susceptible regions and inverted repeats showed detrimental effects on plasmids’ performance as biopharmaceuticals. On the other hand, careful selection of promoter, polyadenylation signal, codon optimization and/or insertion of introns or nuclear-targeting sequences for therapeutic protein expression can enhance the clinical efficacy. Minimal vectors, which are devoid of the bacterial backbone and consist exclusively of the eukaryotic expression cassette, demonstrate better performance in terms of expression levels, bioavailability, transfection rates and increased therapeutic effects. Although the results are promising, minimal vectors have not taken over the conventional plasmids in clinical trials due to challenging manufacturing issues.