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Science 296 (5567): 550-553

Copyright © 2002 by the American Association for the Advancement of Science

A System for Stable Expression of Short Interfering RNAs in Mammalian Cells

Thijn R. Brummelkamp,1 René Bernards,13 Reuven Agami123*

Mammalian genetic approaches to study gene function have been hampered by the lack of tools to generate stable loss-of-function phenotypes efficiently. We report here a new vector system, named pSUPER, which directs the synthesis of small interfering RNAs (siRNAs) in mammalian cells. We show that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Stable expression of siRNAs using this vector mediates persistent suppression of gene expression, allowing the analysis of loss-of-function phenotypes that develop over longer periods of time. Therefore, the pSUPER vector constitutes a new and powerful system to analyze gene function in a variety of mammalian cell types.

1 Division of Molecular Carcinogenesis,
2 Division of Tumor Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, Netherlands.
3 Center for Biomedical Genetics, Netherlands.
*   To whom correspondence should be addressed. E-mail: r.agami{at}

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Silencing of the GnRH Type 1 Receptor Blocks the Antiproliferative Effect of the GnRH Agonist, Leuprolide, on the Androgen-Independent Prostate Cancer Cell Line DU145.
Y. Ko, Y. Ha, J. Kim, S. Kang, H. Jang, S. Kang, H. Park, and J. Cheon (2011)
Journal of International Medical Research 39, 729-739
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