The role of cytoskeleton networks on lipid-mediated delivery of DNA.
BACKGROUND
Lipid-mediated delivery of DNA is hindered by extracellular and intracellular barriers that significantly reduce the transfection efficiency of synthetic nonviral vectors.


RESULTS
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.


CONCLUSIONS
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.
Department of Molecular Medicine, 'Sapienza' University of Rome, Viale Regina Elena, 324, 00161, Rome, Italy.
Stefano
S
Coppola
Francesco
F
Cardarelli
Daniela
D
Pozzi
Laura C
LC
Estrada
Michelle A
MA
Digman
Enrico
E
Gratton
Angelo
A
Bifone
Carlotta
C
Marianecci
Giulio
G
Caracciolo
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D016466:CHO Cells; D002412:Cations; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage* / Q000493:pharmacokinetics; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes; D018613:Microscopy, Confocal; D008870:Microtubules / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D053778:Thiazolidines / Q000494:pharmacology; D014162:Transfection
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002412:Cations; D008055:Lipids; D008081:Liposomes; D053778:Thiazolidines; D004247:DNA; C037068:latrunculin B; D015739:Nocodazole