Gene Silencing in Insect Cells Using RNAi

Hsuan-Chen Wu; John C March; William E Bentley

doi:10.1007/978-1-4939-3043-2_24

Gene Silencing in Insect Cells Using RNAi

Methods Mol Biol. 2016:1350:469-76. doi: 10.1007/978-1-4939-3043-2_24.

Authors

Hsuan-Chen Wu¹, John C March², William E Bentley^{3

4}

Affiliations

¹ Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan.
² Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY, USA.
³ Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA. bentley@umd.edu.
⁴ Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, MD, USA. bentley@umd.edu.

PMID: 26820874
DOI: 10.1007/978-1-4939-3043-2_24

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.

Keywords: Gene silencing; RNA interference; RNAi; SP6; T7; Transcription; dsRNA; mRNA.

MeSH terms

Animals
Cell Line
Genetic Techniques*
Insecta / cytology*
RNA Interference*
RNA, Double-Stranded / genetics
Transfection

Substances

RNA, Double-Stranded