1. The role of cytoskeleton networks on lipid-mediated delivery of DNA
- Author
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Stefano Coppola, Francesco Cardarelli, Giulio Caracciolo, Daniela Pozzi, Laura C. Estrada, Enrico Gratton, Angelo Bifone, Michelle A. Digman, Carlotta Marianecci, Coppola, Stefano, Cardarelli, Francesco, Pozzi, Daniela, Estrada, Laura C., Digman, Michelle A., Gratton, Enrico, Bifone, Angelo, Marianecci, Carlotta, and Caracciolo, Giulio
- Subjects
Cytoplasm ,DRUG DELIVERY ,Pharmaceutical Science ,Arp2/3 complex ,Microtubules ,chemistry.chemical_compound ,Cricetinae ,Thiazolidine ,Cytoskeleton ,Microscopy, Confocal ,Nocodazole ,Transfection ,Lipid ,Lipids ,Cell biology ,Liposome ,Medicina Básica ,Thiazolidines ,Cricetulu ,Intracellular ,CIENCIAS MÉDICAS Y DE LA SALUD ,Genética Humana ,Biological Transport, Active ,Microtubule ,CHO Cells ,INGENIERÍAS Y TECNOLOGÍAS ,Biology ,Article ,Cricetulus ,Cations ,Animals ,Actin ,Nanotecnología ,Cation ,Animal ,DNA ,Bridged Bicyclo Compounds, Heterocyclic ,Nano-materiales ,SINGLE PARTICLE TRACKING ,Actins ,CHO Cell ,chemistry ,Liposomes ,biology.protein - Abstract
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. Conclusion: 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. Fil: Coppola, Stefano. Sapienza University of Rome. Dipartimento di Medicina Molecolare; Italia Fil: Cardarelli, Francesco. Istituto Italiano di Tecnologia. Center for Nanotechnology Innovation at NEST; Italia Fil: Pozzi, Daniela. Sapienza University of Rome. Dipartimento di Medicina Molecolare; Italia Fil: Estrada, Laura Cecilia. University of California. Department of Biomedical Engineering. Laboratory for Fluorescence Dynamics; Estados Unidos. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física. Laboratorio de Electrónica Cuántica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Física de Buenos Aires; Argentina Fil: Digman, Michelle. University of California. Department of Biomedical Engineering. Laboratory for Fluorescence Dynamics; Estados Unidos Fil: Gratton, Enrico. University of California. Department of Biomedical Engineering. Laboratory for Fluorescence Dynamics; Estados Unidos Fil: Bifone, Angelo . Istituto Italiano di Tecnologia. Center for Nanotechnology Innovation at NEST; Italia Fil: Marianecci, Carlotta. Sapienza University of Rome. Dipartimento di Chimica e Tecnologie del Farmaco; Italia Fil: Caracciolo, Giulio. Sapienza University of Rome. Dipartimento di Medicina Molecolare; Italia
- Published
- 2013