Abstract
Lipid-mediated delivery of DNA is hindered by extracellular and intracellular barriers that significantly reduce the transfection efficiency of synthetic nonviral vectors. In this study we investigated the role of the actin and microtubule networks on the uptake and cytoplasmic transport of multicomponent cationic liposome-DNA complexes in CHO-K1 live cells by means of confocal laser scanning microscopy and 3D single particle tracking. Treatment with actin (latrunculin B)- and microtubule-disrupting (nocodazole) reagents indicated that intracellular trafficking of complexes predominantly involves microtubule-dependent active transport. We found that the actin network has a major effect on the initial uptake of complexes, while the microtubule network is mainly responsible for the subsequent active transportation to the lysosomes. Collectively, a strategy to improve the efficiency of lipid gene vectors can be formulated. We could find a lipid formulation that allows the nanoparticles to avoid the microtubule pathway to lysosomes.
Highlights
ObjectivesThe goal of this study was to investigate the role of cytoskeleton structure on the uptake and intracellular dynamics of MC lipoplexes in CHO-K1 live cells
Lipid-mediated delivery of DNA is hindered by extracellular and intracellular barriers that significantly reduce the transfection efficiency of synthetic nonviral vectors
Since intracellular trafficking and endosomal escape are strongly related to each other, novel insights into the cytoplasmic transport of MC lipoplexes could help to explain their unusual tendency to escape from endosomes and to prevent lysosomal degradation
Summary
The goal of this study was to investigate the role of cytoskeleton structure on the uptake and intracellular dynamics of MC lipoplexes in CHO-K1 live cells. The aim of the present work was to gain novel quantitative insights into the role played by both actin microfilaments and microtubules in the intracellular dynamics of MC lipoplexes
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