Methods for Gene Silencing With RNAi

Abstract

This chapter describes a technique for synthesizing and transfecting double stranded RNA (dsRNA) for RNA interference in Sf-21 cell culture. Transfection with dsRNA only requires 1 h and the cells are usually recovered 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 and MEGAscript T7 kit from Ambion and the Superscript III kit from Invitrogen.