A system for stable expression of short interfering RNAs in mammalian cells

TR Brummelkamp, R Bernards, R Agami - science, 2002 - science.org
science, 2002science.org
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 …
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.
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