RNAi Expression Vectors in Mammalian Cells

  • Makoto Miyagishi
  • Kazunari Taira
Part of the Methods in Molecular Biology™ book series (MIMB, volume 252)

Abstract

RNA interference (RNAi) is a recently developed technique for gene silencing by introducing dsRNA into cells, and it is shown to work in mammalian cells when siRNAs are used. Several groups have developed vector-based siRNA expression systems that can induce RNAi in living cells. These vector systems use a pol III promoter, such as U6 or H1, and are classified into two groups based on the form of expressed RNAs: tandem-type and hairpin-type. Here, we describe how to generate these siRNA expression vectors and outline the experimental procedure for suppressing the expression of a reporter gene by transient transfection of a siRNA expression vector.

Keywords

Vortex Agar EDTA Agarose Electrophoresis 

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Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Makoto Miyagishi
    • 1
  • Kazunari Taira
    • 1
  1. 1.Department of Chemistry and Biotechnology, School of EngineeringThe University of TokyoTokyoJapan

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