title: Simple and rapid synthesis of siRNA derived from in vitro transcribed shRNA.
abstract: 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.
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D034741:RNA, Small Interfering / Q000138:chemical synthesis*; D014158:Transcription, Genetic; D014764:Viral Proteins
chemical_list: D034741:RNA, Small Interfering; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases