Scientific article
Open access

RNA interference in mammalian cells using siRNAs synthesized with T7 RNA polymerase

Published inNucleic acids research, vol. 30, no. 10, e46
Publication date2002

Methods that allow the specific silencing of a desired gene are invaluable tools for research. One of these is based on RNA interference (RNAi), a process by which double-stranded RNA (dsRNA) specifically suppresses the expression of a target mRNA. Recently, it has been reported that RNAi also works in mammalian cells if small interfering RNAs (siRNAs) are used to avoid activation of the interferon system by long dsRNA. Thus, RNAi could become a major tool for reverse genetics in mammalian systems. However, the high cost and the limited availability of the short synthetic RNAs and the lack of certainty that a designed siRNA will work present major drawbacks of the siRNA technology. Here we present an alternative method to obtain cheap and large amounts of siRNAs using T7 RNA polymerase. With multiple transfection procedures, including calcium phosphate co-precipitation, we demonstrate silencing of both exogenous and endogenous genes.

  • Cell Line
  • Cloning, Molecular/methods
  • DNA-Directed RNA Polymerases/metabolism
  • Gene Silencing
  • Green Fluorescent Proteins
  • Hela Cells
  • Humans
  • Luminescent Proteins/genetics/metabolism
  • Plasmids/genetics
  • RNA, Double-Stranded/genetics/metabolism
  • RNA, Messenger/genetics/metabolism
  • Recombinant Fusion Proteins/genetics/metabolism
  • Transfection
  • Viral Proteins
Citation (ISO format)
DONZE, Olivier, PICARD, Didier. RNA interference in mammalian cells using siRNAs synthesized with T7 RNA polymerase. In: Nucleic acids research, 2002, vol. 30, n° 10, p. e46.
Main files (1)
Article (Published version)
ISSN of the journal0305-1048

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