Micrometric DNA/PEI polyplexes correlate with higher transient gene expression yields in HEK 293 cells

Irene González-Domínguez*, Eduard Puente-Massaguer, Jesús Lavado-García, Laura Cervera, Francesc Gòdia

*Autor corresponent d’aquest treball

Producció científica: Contribució a revistaArticleRecercaAvaluat per experts

13 Cites (Scopus)

Resum

DNA delivery with polyethylenimine (PEI) has been widely used in the last three decades for the transfection of mammalian cells. Advances in novel characterization techniques at the nanometric scale offer new opportunities to revisit the physicochemical properties of DNA/PEI polyplexes that lead to efficient transfection. In this work, these properties are tuned by studying the synergies between simple parameters such as NaCl concentration, pH and incubation time in the DNA/PEI polyplex preparation protocol by means of Design of Experiments (DoE). By doing so, a model is obtained where an optimal NaCl concentration of 125 mM and an incubation time of 11 min provided the highest transfection yields. Correlation analyses between the physicochemical properties of DNA/PEI polyplexes and the predicted model responses revealed the existence of an optimal degree of aggregation in the pre-complexing solution to attain the highest transfection efficiencies. The presence of these micrometric DNA/PEI polyplex aggregates was confirmed by several nanoparticle characterization techniques including cryo-TEM, DLS and flow virometry. The findings provide a better understanding of the role of DNA/PEI aggregates in transient gene expression approaches, in particular considering that similar complexation protocols and saline solutions are widely used for the transfection of mammalian cell cultures.

Idioma originalAnglès
Pàgines (de-a)87-96
Nombre de pàgines10
RevistaNew Biotechnology
Volum68
DOIs
Estat de la publicacióPublicada - 25 de maig 2022

Fingerprint

Navegar pels temes de recerca de 'Micrometric DNA/PEI polyplexes correlate with higher transient gene expression yields in HEK 293 cells'. Junts formen un fingerprint únic.

Com citar-ho