Protein inhibition by microinjection and RNA-mediated interference in tissue culture cells

complementary approaches to study protein function.

Jane R. Stout, Rania S. Rizk, Claire Walczak

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A major goal in cell biology is to understand the molecular mechanisms of the biological process under study, which requires functional information about the roles of individual proteins in the cell. For many non-genetic model organisms researchers have relied on the use of inhibitory reagents, such as antibodies that can be microinjected into cells. More recently, the advent of RNA-mediated interference (RNAi) has allowed scientists to knockdown individual proteins and to examine the consequences of the knockdown. In this chapter we present a comparison between microinjection of inhibitory reagents and RNAi for the analysis of protein function in mammalian tissue culture cells, providing both a description of the techniques as well as a discussion of the benefits and drawbacks of each approach. In addition, we present a strategy to employ RNAi for organisms without a sequenced genome. While the focus of our research is on the organization of the mitotic spindle during cell division and thus the examples utilized are from that system, the approaches described here should be readily applicable to multiple experimental models.

Original languageEnglish
Pages (from-to)77-97
Number of pages21
JournalMethods in molecular biology (Clifton, N.J.)
Volume518
StatePublished - 2009

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Microinjections
RNA Interference
Cell Culture Techniques
Biological Phenomena
Spindle Apparatus
Proteins
Cell Division
Cell Biology
Theoretical Models
Research Personnel
Genome
Antibodies
Research

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

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