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Which mechanism for nuclear import of plasmid DNA complexed with polyethylenimine derivatives?
Author(s) -
Grosse Stéphanie,
Thévenot Guiti,
Monsigny Michel,
Fajac Isabelle
Publication year - 2006
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.915
Subject(s) - polyethylenimine , transfection , plasmid , nuclear localization sequence , nuclear transport , biology , dna , microbiology and biotechnology , mitosis , nuclear dna , cell nucleus , gene , biochemistry , mitochondrial dna
Background To investigate the nuclear import mechanism of plasmid/polyethylenimine (PEI) derivative complexes and the putative nuclear targeting of therapeutic genes by the use of oligosaccharides, we have studied the nuclear import of plasmid DNA complexed either with PEI or with lactosylated PEI (Lac–PEI) in cystic fibrosis human airway epithelial cells (∑CFTE29o‐ cells). Methods and results Cells were synchronized by a double‐thymidine block protocol and gene transfer efficiency was evaluated: Lac–PEI‐ and PEI‐mediated gene transfer was greatly increased when cells have undergone mitosis during the course of transfection. However, both types of complexes were able to transfect some growth‐arrested cells. When the nuclear import of plasmid/Lac–PEI or plasmid/unsubstituted PEI complexes was studied in digitonin‐permeabilized cells, the nuclear uptake of both types of complexes did not follow the classic pathway of nuclear localization sequence (NLS)‐containing proteins and lactose residues did not act as a nuclear localization signal. Conclusions Our results show that for complexes made with PEI derivatives, the major route for plasmid DNA nuclear entry is a passive nuclear importation during mitosis when the nuclear membrane temporarily breaks down. However, albeit to a lesser extent as that observed in dividing cells, a plasmid DNA importation also occurs in nondividing cells by a yet unknown mechanism. Copyright © 2006 John Wiley & Sons, Ltd.