title: Gene Silencing in Insect Cells Using RNAi.
abstract: A technique is described for synthesizing and transfecting double stranded RNA (dsRNA) for RNA interference (RNAi) in Sf-21 cell culture. Transfection with dsRNA only requires an hour and the cells usually recover within 12 h. Suggestions for designing dsRNA are included in the methods. Furthermore, websites are provided for rapid and effective dsRNA design. Three kits are essential for using the described methods: RNAqueous®-4PCR, Megascript™ T7 kit, and the Superscript™ III kit from Life Technologies, Inc.
authors.affiliation: Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan.
authors.forename: Hsuan-Chen
authors.initials: HC
authors.lastname: Wu
authors.affiliation: Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: March
authors.affiliation: Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA. bentley@umd.edu.|Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, MD, USA. bentley@umd.edu.
authors.forename: William E
authors.initials: WE
authors.lastname: Bentley
mesh_terms: D000818:Animals; D002460:Cell Line; D005821:Genetic Techniques; D007313:Insecta / Q000166:cytology*; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D014162:Transfection
chemical_list: D012330:RNA, Double-Stranded
keywords: Gene silencing; RNA interference; RNAi; SP6; T7; Transcription; dsRNA; mRNA