Intracellular trafficking of cationic liposome–DNA complexes in living cells

Stefano Coppola; Laura C. Estrada; Michelle A. Digman; Daniela Pozzi; Francesco Cardarelli; Enrico Gratton; Giulio Caracciolo

doi:10.1039/C2SM25532D

Intracellular trafficking of cationic liposome–DNA complexes in living cells†

Stefano Coppola,^a Laura C. Estrada,^b Michelle A. Digman,^b Daniela Pozzi,^a Francesco Cardarelli,^c Enrico Gratton^b and Giulio Caracciolo*^a

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* Corresponding authors

^a Department of Molecular Medicine, “Sapienza” University of Rome, Viale Regina Elena 324, Rome, Italy
E-mail: giulio.caracciolo@uniroma1.it

^b Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, 3120 Natural Sciences 2, Irvine, California 92697-2715, USA

^c Center for Nanotechnology Innovation, @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127 Pisa, Italy

Abstract

Three-dimensional single-particle tracking (SPT) was used to calculate the mean square displacement (MSD) and the diffusion coefficients of multicomponent cationic liposome–DNA complexes (lipoplexes) in CHO-K1 living cells. In untreated (NT) control cells, we found that the intracellular lipoplex motion was either directed or Brownian with active transportation being definitely more frequent (more than 70%) than Brownian diffusion. The MSD analysis was supported by the calculation of the three-dimensional asphericity, A₃, which was close to unity, denoting the preponderant occurrence of movement along a direction. To elucidate the role of the cytoskeleton structure in the lipoplex trafficking, cells were treated with cytoskeleton (actin microfilaments and microtubules) polymerization inhibitors (latrunculin B and nocodazole, respectively). When cells were treated with inhibitors, the lipoplex movement tended towards a random walk at the expense of directed motion. The disassembly of microtubules had a stronger effect on the reduction of directional movement than that of actin microfilaments. Relevance of the results for enhanced gene delivery is discussed.

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DOI: https://doi.org/10.1039/C2SM25532D
Article type: Paper
Submitted: 06 Mar 2012
Accepted: 01 Jun 2012
First published: 02 Jul 2012

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Soft Matter, 2012,8, 7919-7927

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Intracellular trafficking of cationic liposome–DNA complexes in living cells

S. Coppola, L. C. Estrada, M. A. Digman, D. Pozzi, F. Cardarelli, E. Gratton and G. Caracciolo, Soft Matter, 2012, 8, 7919 DOI: 10.1039/C2SM25532D

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Soft Matter

Intracellular trafficking of cationic liposome–DNA complexes in living cells†

Abstract

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Intracellular trafficking of cationic liposome–DNA complexes in living cells

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