Access the full text.
Sign up today, get DeepDyve free for 14 days.
I. Mortimer, P. Tam, I. Maclachlan, R. Graham, E. Saravolac, P. Joshi (1999)
Cationic lipid-mediated transfection of cells in culture requires mitotic activityGene Therapy, 6
M. James, T. Giorgio (2000)
Nuclear-associated plasmid, but not cell-associated plasmid, is correlated with transgene expression in cultured mammalian cells.Molecular therapy : the journal of the American Society of Gene Therapy, 1 4
F. Soubrier, B. Cameron, B. Manse, S. Somarriba, C. Dubertret, G. Jaslin, G. Jung, C. Caer, D. Dang, J. Mouvault, D. Scherman, J. Mayaux, J. Crouzet (1999)
pCOR: a new design of plasmid vectors for nonviral gene therapyGene Therapy, 6
V. Escriou, Carole Ciolina, Florence Lacroix, G. Byk, D. Scherman, P. Wils (1998)
Cationic lipid-mediated gene transfer: effect of serum on cellular uptake and intracellular fate of lipopolyamine/DNA complexes.Biochimica et biophysica acta, 1368 2
D. Stoffler, B. Fahrenkrog, U. Aebi (1999)
The nuclear pore complex: from molecular architecture to functional dynamics.Current opinion in cell biology, 11 3
S. Adam (1999)
Transport pathways of macromolecules between the nucleus and the cytoplasm.Current opinion in cell biology, 11 3
Yuhong Xu, F. Szoka (1996)
Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection.Biochemistry, 35 18
Carole Ciolina, G. Byk, Francis Blanche, Vincent Thuillier, D. Scherman, P. Wils (1999)
Coupling of nuclear localization signals to plasmid DNA and specific interaction of the conjugates with importin alpha.Bioconjugate chemistry, 10 1
G. Pedrali-Noy, S. Spadari, A. Miller-Faurès, A. Miller, J. Kruppa, G. Koch (1980)
Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.Nucleic acids research, 8 2
A. Fasbender, J. Zabner, BG Zeiher, M. Welsh (1997)
A low rate of cell proliferation and reduced DNA uptake limit cationic lipid-mediated gene transfer to primary cultures of ciliated human airway epitheliaGene Therapy, 4
D. Lechardeur, Sohn Kj, Martin Haardt, Phalgun Joshi, M. Monck, Roger Graham, B. Beatty, Jeremy Squire, Hugh O'Brodovich, G. Lukács (1999)
Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transferGene Therapy, 6
Martin Dowty, P. Williams, Guofeng Zhang, J. Hagstrom, Jon Wolff (1995)
Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.Proceedings of the National Academy of Sciences of the United States of America, 92 10
M. Subramanian, S. Diamond, H. Ma, Shin Nakamura, Satoko Mitsunaga, Keiko Shimizu, Makoto Hirano, Toshitaka Imamura, Akihiro Abe, N. Emi (1999)
Nonclassical Nuclear Localization Signal Provides for High Efficiency, Nonviral Gene Transfer to Nondividing EndotheliumNature Biotechnology, 17 Suppl 4
Non viral
L. Brandén, A. Mohamed, C. Smith (1999)
A peptide nucleic acid–nuclear localization signal fusion that mediates nuclear transport of DNANature Biotechnology, 17
Martina Wilke, E. Fortunati, M. Broek, A. Hoogeveen, B. Scholte (1996)
Efficacy of a peptide-based gene delivery system depends on mitotic activity.Gene therapy, 3 12
M. Sebestyén, J. Ludtke, M. Bassik, Guofeng Zhang, V. Budker, E. Lukhtanov, J. Hagstrom, J. Wolff (1998)
DNA vector chemistry: The covalent attachment of signal peptides to plasmid DNANature Biotechnology, 16
(1994)
Synchronization of normal diploid and transformed mammalian cells
C. Neves, G. Byk, D. Scherman, P. Wils (1999)
Coupling of a targeting peptide to plasmid DNA by covalent triple helix formationFEBS Letters, 453
W. Zauner, S. Brunner, Michael Buschle, M. Ogris, Ernst Wagner, Ernst Wagner (1999)
Differential behaviour of lipid based and polycation based gene transfer systems in transfecting primary human fibroblasts: a potential role of polylysine in nuclear transport.Biochimica et biophysica acta, 1428 1
Gary Whittaker, Ari Helenius (1998)
Nuclear import and export of viruses and virus genomes.Virology, 246 1
Wen-Chi Tseng, F. Haselton, Todd Giorgio (1999)
Mitosis enhances transgene expression of plasmid delivered by cationic liposomes.Biochimica et biophysica acta, 1445 1
M. Capecchi (1980)
High efficiency transformation by direct microinjection of DNA into cultured mammalian cellsCell, 22
S. Adam, Rachel Marr, L. Gerace (1990)
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factorsThe Journal of Cell Biology, 111
M. Brisson, W. Tseng, C. Almonte, S. Watkins, L. Huang (1999)
Subcellular trafficking of the cytoplasmic expression system.Human gene therapy, 10 16
J. Zabner, A. Fasbender, T. Moninger, K. Poellinger, M. Welsh (1995)
Cellular and Molecular Barriers to Gene Transfer by a Cationic Lipid (*)The Journal of Biological Chemistry, 270
C. Kitson, B. Angel, D. Judd, S. Rothery, N. Severs, A. Dewar, L. Huang, S. Wadsworth, S. Cheng, D. Geddes, E. Alton (1999)
The extra- and intracellular barriers to lipid and adenovirus-mediated pulmonary gene transfer in native sheep airway epitheliumGene Therapy, 6
Yang Jp, Leaf Huang (1997)
Overcoming the inhibitory effect of serum on lipofection by increasing the charge ratio of cationic liposome to DNAGene Therapy, 4
Song Li, Leaf Huang (2000)
Nonviral gene therapy: promises and challengesGene Therapy, 7
Hélène Pollard, J. Remy, G. Loussouarn, S. Demolombe, J. Behr, D. Escande (1998)
Polyethylenimine but Not Cationic Lipids Promotes Transgene Delivery to the Nucleus in Mammalian Cells*The Journal of Biological Chemistry, 273
A. Mountain (2000)
Gene therapy: the first decade.Trends in biotechnology, 18 3
S. Brunner, T. Sauer, S. Carotta, M. Cotten, M. Saltik, E. Wagner (2000)
Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirusGene Therapy, 7
A. Subramanian, P. Ranganathan, S. Diamond (1999)
Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cellsNature Biotechnology, 17
M. Zanta, P. Belguise-Valladier, J. Behr (1999)
Gene delivery: a single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus.Proceedings of the National Academy of Sciences of the United States of America, 96 1
H. Farhood, N. Serbina, Leaf Huang (1995)
The role of dioleoyl phosphatidylethanolamine in cationic liposome mediated gene transfer.Biochimica et biophysica acta, 1235 2
Carole Neves, G. Byk, V. Escriou, Florence Bussone, D. Scherman, P. Wils (2000)
Novel method for covalent fluorescent labeling of plasmid DNA that maintains structural integrity of the plasmid.Bioconjugate chemistry, 11 1
Background Cationic lipid‐mediated gene transfer is a promising approach for gene therapy. However, despite the significant amount of lipoplexes internalized by target cells, transgene expression remains too low. Obstacles to nuclear accumulation of plasmid DNA include: the passage of DNA across the cellular membrane, the dismantling of nucleolipidic particles in the cytoplasm and the nuclear import of plasmid DNA. The purpose of the present study was to evaluate the impact of cell status on cationic lipid‐mediated transfer. Methods Cells were either growth‐arrested (by aphidicolin) or synchronized (by a classical double‐thymidine block protocol) and cationic lipid‐mediated transfection of these cells was evaluated. For the study of the nuclear import of plasmid DNA, two techniques were developed: microinjection of plasmid DNA into intact cells, and the use of cells permeabilized with digitonin. Results When CV‐1 cells were growth‐arrested by aphidicolin, cationic lipid‐mediated gene transfer was inhibited. Hela cells were synchronized and incubated with lipoplexes at different times after release of the block. Gene expression was greatly enhanced when cells underwent mitosis. When transfection was performed during the early period after block release, when fewer than 5% of the cells had divided, gene expression was carefully quantified and could be attributed to cells that escaped cell cycle block. However, by direct analysis of nuclear import of GFP‐coding plasmid using cytoplasmic microinjection, GFP expression could be detected in a few cells that had not divided. Conclusions Cationic lipid‐mediated gene transfer efficiency increased when cells underwent mitosis. However, when cells did not divide, gene transfer was not completely abolished. Nuclear import of plasmid was greatly facilitated by a mitotic event. In non‐mitotic cells, nuclear envelope crossing by plasmid DNA could be detected but was a very rare event. Copyright © 2001 John Wiley & Sons, Ltd.
Journal of Gene Medicine – Wiley
Published: Mar 1, 2001
Keywords: cationic lipid; nuclear import; plasmid DNA; synchronization; cell cycle
Read and print from thousands of top scholarly journals.
Already have an account? Log in
Bookmark this article. You can see your Bookmarks on your DeepDyve Library.
To save an article, log in first, or sign up for a DeepDyve account if you don’t already have one.
Copy and paste the desired citation format or use the link below to download a file formatted for EndNote
Access the full text.
Sign up today, get DeepDyve free for 14 days.
All DeepDyve websites use cookies to improve your online experience. They were placed on your computer when you launched this website. You can change your cookie settings through your browser.