Simple and rapid synthesis of siRNA derived from in vitro transcribed shRNA.
Nucleic Acids Res Suppl. 2003;(3):249-50
Katoh T, Susa M, Suzuki T, Umeda N, Watanabe K, Suzuki T.

Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg. FSB-301, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562, Japan.

Temporal gene silencing in mammalian cells using small interfering RNA (siRNA) is an invaluable tool for mammalian genetics and is becoming established. However, systematic studies of siRNA such as large-scale target validations are limited due to the high cost of chemical synthesis of double-stranded RNAs. Here, we devise a simple, rapid, practical and cost-effective method for preparing active siRNA derived from short hairpin (sh) RNA which is transcribed from a single-stranded synthetic DNA template using T7 RNA polymerase. This method doesn't require any sequence-limitation in the selection of the target region of genes. We demonstrate efficient silencing of several genes by the transcribed siRNAs obtained by this method.

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