Protocol

Neurodegeneration

Volume 793 of the series Methods in Molecular Biology pp 457-477

Date:

Specific Gene Silencing Using RNAi in Cell Culture

  • Chunxing YangAffiliated withDepartment of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School
  • , Linghua QiuAffiliated withDepartment of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School
  • , Zuoshang XuAffiliated withDepartment of Biochemistry and Molecular Pharmacology, Cell Biology, Neuroscience Program, University of Massachusetts Medical School Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

RNA interference (RNAi) is a conserved cellular mechanism in most eukaryotes that can mediate specific gene silencing. Since its discovery in 1998, rapid progress has been made in understanding its basic mechanism and its application in research and drug discovery. In recent years, the application of RNAi in research, including research in neurodegeneration, has expanded rapidly such that it has become a regular tool for reverse genetics in cultured cells in many labs. However, an incomplete understanding of the RNAi mechanism and worries about its pitfalls still intimidate many others. Here, we present a streamlined and simple protocol for the design and implementation of an RNAi experiment in cultured cells, aiming to enable those who are inexperienced with RNAi to apply this powerful method in their research, particularly in the field of neurodegeneration.

Key words

RNAi shRNA siRNA miRNA Gene silencing Knockdown