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Cell cycle dependent transcription, a determinant factor of heterogeneity in cationic lipid-mediated transgene expression.

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Title: Cell cycle dependent transcription, a determinant factor of heterogeneity in cationic lipid-mediated transgene expression.
Authors: Akita, Hidetaka Browse this author
Ito, Rie Browse this author
Kamiya, Hiroyuki Browse this author
Kogure, Kentaro Browse this author
Harashima, Hideyoshi Browse this author →KAKEN DB
Keywords: heterogeneity
nuclear delivery
gene transfer
Issue Date: Mar-2007
Publisher: John Wiley & Sons
Journal Title: The Journal of Gene Medicine
Volume: 9
Issue: 3
Start Page: 197
End Page: 207
Publisher DOI: 10.1002/jgm.1010
PMID: 17351985
Abstract: Background: Heterogeneity of transgene expression, the presence or absence (below the limit of detection) of transgene expression on a cell-by-cell basis, is a severe disadvantage in the use of cationic lipid-mediated gene vectors for gene therapy and experiments in molecular biology. Understandings of intracellular trafficking and the function (transgene expression) of vectors related to cellular physiology are essential in terms of clarifying the mechanism underlying the heterogeneity. Methods: To distinguish the contribution of nuclear transfer efficiency and subsequent intranuclear transcription efficiency to the overall heterogeneity in transgene expression, a novel imaging system was established for the dual visualization of the nuclear transfer of pDNA and marker gene expression (lacZ) in single cells. Results: The expression of LacZ occurred in only approximately 30% of HeLa cells of the nuclear pDNA-positive cells, indicating that intranuclear transcription efficiency contributed to the heterogeneity. Dual imaging against synchronized cells further revealed that the efficiency of nuclear delivery was comparable irrespective of cell cycle status, which is contrary to the generally accepted hypothesis that nuclear import of pDNA is enhanced during cell division when the nuclear membrane structure is perturbed. The most significant finding in the present study is that nuclear transcription efficiency in terms of the ratio of LacZ-positive cells to nuclear pDNA-positive cells drastically increased in the late S and G2/M phase. Conclusions: This is the first demonstration to show that cell cycle dependent intranuclear transcription appears to be responsible for the overall heterogeneity of transgene expression. Copyright © 2007 John Wiley & Sons, Ltd.
Rights: Copyright © 2007 John Wiley & Sons, Inc., The Journal of Gene Medicine, Vol. 9, issue 3, pp. 197-207
Type: article (author version)
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 原島 秀吉

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