Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro

Zsuzsa Bebök, Anna M. Abai, Jian Yun Dong, Scott A. King, Kevin L. Kirk, Gabor Berta, Brian W. Hughes, Andrew Kraft, Stephen W. Burgess, Walter Shaw, Philip L. Felgner, Eric J. Sorscher

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Cationic liposome-mediated gene transfer has become increasingly important in the development of experimental therapies for human diseases, such as melanoma, human immunodeficiency virus infection, cystic fibrosis and alpha-1 antitrypsin deficiency. However, very little is known about the mechanisms by which lipid-mediated gene transfer occurs. We studied the kinetics of plasmid delivery and expression by using this technique. Plasmid entry in the cystic fibrosis respiratory epithelial cell line 2CFSME0-, as well as in two other cell lines (HeP 2g and HeLa) occurred in 95 to 100% of cells within 1 hr of the initiation of lipid-mediated gene transfer. In hepatic and respiratory cells, transcription of a construct containing the cystic fibrosis transmembrane conductance regulator was observed in more than 80% of the cell population; similarly high levels of plasmid utilization were obtained in studies of HLA-B7 expression in human melanoma cells. Studies directly relevant to current human trials of lipid-mediated gene transfer indicate that plasmid entry, transcription and translation are often surprisingly efficient, and may occur in nearly 100% of human cells in culture when sensitive methods for detection are used. Furthermore, conventional X-gal immunohistochemistry markedly underestimates transfection efficiency during transient gene expression. These studies point to a new mechanistic understanding of the features that limit expression by using cationic liposomes.

Original languageEnglish (US)
Pages (from-to)1462-1469
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume279
Issue number3
StatePublished - Dec 1996
Externally publishedYes

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Plasmids
Lipids
Cystic Fibrosis
Liposomes
Genes
Melanoma
HLA-B7 Antigen
alpha 1-Antitrypsin Deficiency
Cell Line
Cystic Fibrosis Transmembrane Conductance Regulator
Investigational Therapies
Virus Diseases
Transfection
Hepatocytes
Cell Culture Techniques
Epithelial Cells
Immunohistochemistry
HIV
Gene Expression
In Vitro Techniques

ASJC Scopus subject areas

  • Pharmacology

Cite this

Bebök, Z., Abai, A. M., Dong, J. Y., King, S. A., Kirk, K. L., Berta, G., ... Sorscher, E. J. (1996). Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro. Journal of Pharmacology and Experimental Therapeutics, 279(3), 1462-1469.

Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro. / Bebök, Zsuzsa; Abai, Anna M.; Dong, Jian Yun; King, Scott A.; Kirk, Kevin L.; Berta, Gabor; Hughes, Brian W.; Kraft, Andrew; Burgess, Stephen W.; Shaw, Walter; Felgner, Philip L.; Sorscher, Eric J.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 279, No. 3, 12.1996, p. 1462-1469.

Research output: Contribution to journalArticle

Bebök, Z, Abai, AM, Dong, JY, King, SA, Kirk, KL, Berta, G, Hughes, BW, Kraft, A, Burgess, SW, Shaw, W, Felgner, PL & Sorscher, EJ 1996, 'Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro', Journal of Pharmacology and Experimental Therapeutics, vol. 279, no. 3, pp. 1462-1469.
Bebök, Zsuzsa ; Abai, Anna M. ; Dong, Jian Yun ; King, Scott A. ; Kirk, Kevin L. ; Berta, Gabor ; Hughes, Brian W. ; Kraft, Andrew ; Burgess, Stephen W. ; Shaw, Walter ; Felgner, Philip L. ; Sorscher, Eric J. / Efficiency of plasmid delivery and expression after lipid-mediated gene transfer to human cells in vitro. In: Journal of Pharmacology and Experimental Therapeutics. 1996 ; Vol. 279, No. 3. pp. 1462-1469.
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