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The apparent uptake of fluorescently labeled siRNAs by electroporated cells depends on the fluorochrome

Lookup NU author(s): Professor Olaf Heidenreich

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Abstract

Transfection of mammalian cells with preformed small interfering RNAs (siRNAs) permits a transient and often specific reduction of gene expression. It is possible to rapidly examine the uptake of siRNAs by transfection with fluorescently labeled siRNAs. We examined the apparent uptake of such siRNAs by several leukemic cell lines after electroporation. We show that Cy3 and Cy5-labeled siRNAs cause a significant amount of cell fluorescence, as judged by flow cytometry. In contrast, several fluorescein-labeled siRNAs could not be detected. Nevertheless, such fluoresceinated siRNAs efficiently suppressed a leukemic target gene, demonstrating that siRNA uptake must have taken place. Therefore, for cell electroporation, fluorescein-labeled siRNAs may lead to false negative results and should not be used to examine electroporation-mediated siRNA uptake.

Publication metadata

Author(s): Dunne J, Drescher B, Riehle H, Hadwiger P, Young BD, Krauter J, Heidenreich O

Publication type: Article

Publication status: Published

Journal: Oligonucleotides

Year: 2003

Volume: 13

Issue: 5

Pages: 375-380

ISSN (print): 1545-4576

ISSN (electronic): 1557-8526

URL: http://dx.doi.org/10.1089/154545703322617050

DOI: 10.1089/154545703322617050

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