Document Detail


Which mechanism for nuclear import of plasmid DNA complexed with polyethylenimine derivatives?
MedLine Citation:
PMID:  16685744     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 ( summation operatorCFTE29o- 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.
Authors:
Stéphanie Grosse; Guiti Thévenot; Michel Monsigny; Isabelle Fajac
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The journal of gene medicine     Volume:  8     ISSN:  1099-498X     ISO Abbreviation:  J Gene Med     Publication Date:  2006 Jul 
Date Detail:
Created Date:  2006-07-03     Completed Date:  2006-09-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9815764     Medline TA:  J Gene Med     Country:  England    
Other Details:
Languages:  eng     Pagination:  845-51     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2006 John Wiley & Sons, Ltd.
Affiliation:
Université Paris-Descartes, Faculté de Médecine, AP-HP, Hôpital Cochin, EA 2511, Paris, France.
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MeSH Terms
Descriptor/Qualifier:
Active Transport, Cell Nucleus
Cell Cycle
Cell Line
DNA, Recombinant / administration & dosage,  chemistry,  genetics,  pharmacokinetics*
Gene Transfer Techniques*
Green Fluorescent Proteins / genetics
Humans
Macromolecular Substances
Microinjections
Mitosis
Plasmids / administration & dosage*,  chemistry,  genetics,  pharmacokinetics*
Polyethyleneimine
Recombinant Proteins / genetics
Chemical
Reg. No./Substance:
0/DNA, Recombinant; 0/Macromolecular Substances; 0/Recombinant Proteins; 147336-22-9/Green Fluorescent Proteins; 9002-98-6/Polyethyleneimine

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