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Metafectene is superior to lipofectamine in the transfection of G(s) alpha prostate cancer cells. Mol Biotechnol 2004 Oct;28(2):97-103

Date

10/13/2004

Pubmed ID

15477648

DOI

10.1385/MB:28:2:097

Scopus ID

2-s2.0-21344457489   7 Citations

Abstract

Transfection efficiency of the novel reagent metafectene has not been compared with that of lipofectamine in the published English literature. We used these agents to transfect two prostate cancer cell lines, PC3 and G(s) alpha, with a deoxyribonucleic acid (DNA) expression vector that generates double-stranded ribonucleic acid (RNA) for RNA interference (RNAi). Cotransfection of the green fluorescent protein (GFP) reporter gene revealed that the mean (+/- standard deviation) transfection efficiencies with lipofectamine were 5.8+/-0.4% for PC3 cells and 3.6+/-1.5% for G(s) alpha cells. Mean transfection efficiency with metafectene declined to 0.1+/-0% for PC3 cells but improved to 54.6+/-5.5% for G(s) alpha cells. With G(s) alpha cells, metafectene transfection of GFP plasmid alone yielded 46.9% positive cells, and cotransfection with CD44v9 expression vector yielded 45.9% positive cells. The visual impact of the transfected RNAi construct was detectable at the protein level 4 to 6 d posttransfection and was more dramatic after using metafectene than after using lipofectamine. Thus, in vitro, metafectene transfection efficiency was sufficient to allow us to assess the functional significance of our RNAi construct, suggesting metafectene as an excellent candidate for RNAi-mediated anticancer gene therapy.

Author List

Iczkowski KA, Omara-Opyene AL, Klösel R

Author

Kenneth A. Iczkowski MD Professor in the Pathology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Biotechnology
Blotting, Western
Cell Line, Tumor
DNA
Drug Carriers
GTP-Binding Protein alpha Subunits
Genetic Therapy
Genetic Vectors
Green Fluorescent Proteins
Humans
Lipids
Male
Models, Genetic
Plasmids
Prostatic Neoplasms
RNA Interference
RNA, Double-Stranded
Time Factors
Transfection
jenkins-FCD Prod-398 336d56a365602aa89dcc112f077233607d6a5abc