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A System for Stable Expression of Short Interfering RNAs in Mammalian Cells
Thijn R. Brummelkamp,1René Bernards,13Reuven Agami123*
Mammalian genetic approaches to study gene function have
been hampered by the lack of tools to generate stable loss-of-functionphenotypes efficiently. We report here a new vector system, namedpSUPER, which directs the synthesis of small interfering RNAs(siRNAs)
in mammalian cells. We show that siRNA expression mediatedby this
vector causes efficient and specific down-regulation ofgene
expression, resulting in functional inactivation of the targetedgenes.
Stable expression of siRNAs using this vector mediatespersistent
suppression of gene expression, allowing the analysisof
loss-of-function phenotypes that develop over longer periodsof time.
Therefore, the pSUPER vector constitutes a new and powerfulsystem to
analyze gene function in a variety of mammalian celltypes.
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}nki.nl
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Combined functional and molecular analysis of tumor cell signaling defines 2 distinct myeloma subgroups: Akt-dependent and Akt-independent multiple myeloma.
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Blood
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Megakaryocyte endomitosis is a failure of late cytokinesis related to defects in the contractile ring and Rho/Rock signaling.
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Blood
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|Abstract »|Full Text »|PDF »
Telomere dysfunction and cell survival: roles for distinct TIN2-containing complexes.
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Generation of a highly inducible Gal4->Fluc universal reporter mouse for in vivo bioluminescence imaging.
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|Abstract »|Full Text »|PDF »
Clathrin Assembly Protein AP180 and CALM Differentially Control Axogenesis and Dendrite Outgrowth in Embryonic Hippocampal Neurons.
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