Specific Gene Silencing Using RNAi in Cell Culture

Part of the Methods in Molecular Biology book series (MIMB, volume 793)

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 

Notes

Acknowledgments

This work has been supported by RO1 NS048145 and R21 NS062230-01.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular PharmacologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  2. 2.Department of Biochemistry and Molecular Pharmacology, Cell Biology, Neuroscience ProgramUniversity of Massachusetts Medical SchoolWorcesterUSA

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