title: The actin cytoskeleton has an active role in the electrotransfer of plasmid DNA in mammalian cells.
abstract: Electrotransfer of molecules is a well established technique which finds extensive use for gene transfer and holds great promise for anticancer treatment. Despite its widespread application, the mechanisms governing the entry of DNA into the cell and its intracellular trafficking are not yet known. The aim of this study is to unravel the role of the actin cytoskeleton during gene electrotransfer in cells. We performed single-cell level approaches to observe the organization of the actin cytoskeleton in Chinese hamster ovary (CHO) cells. In addition, we performed experiments at the multiple-cell level to evaluate the efficiency of DNA transfer after alteration of the actin cytoskeleton using the drug latrunculin B. Actin patches colocalizing with the DNA at the plasma membrane were observed with additional characteristics similar to those of the DNA aggregates in terms of time, number, and size. The disruption of the microfilaments reduces the DNA accumulation at the plasma membrane and the gene expression. This is the first direct experimental evidence of the participation of the actin cytoskeleton in DNA electrotransfer.
authors.affiliation: Department of Structural Biology and Biophysics, CNRS, Institut de Pharmacologie et de Biologie Structurale, Toulouse, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Rosazza
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Escoffre
authors.forename: Andreas
authors.initials: A
authors.lastname: Zumbusch
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Rols
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D003599:Cytoskeleton / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004560:Electricity / Q000009:adverse effects; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D010957:Plasmids; D021381:Protein Transport; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D053778:Thiazolidines; D004247:DNA; C037068:latrunculin B