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Transgenic RNA Interference to Investigate Gene Function in the Mouse

  • Tilo Kunath
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 461)

1. Introduction

The necessity of a gene for a particular biological function is often investigated by loss-of-function studies. Although this may be accomplished by a number of means, the most common and def initive strategy is to physically disrupt the genetic locus in a way that results in the production of a nonfunctional protein or prevents transcription, without affecting the activity of nearby genes. In the mouse, this can be performed in several ways. Chemical mutagenesis and gene trap approaches have been very useful to obtain randomly induced mutations (1,2), while homologous recombination in embryonic stem (ES) cells has been the traditional method to disrupt genes in a directed manner (3,4).

A less definitive, but often effective and rapid method to investigate the necessity of a protein-encoding gene is by posttranscriptional suppression. Several experimental strategies exist that either promote mRNA degradation or inhibit translation, including antisense, ribozyme, and...

Keywords

Embryonic Stem Cell Embryonic Stem Cell Line Sense Strand Antisense Strand Embryonic Stem Cell Clone 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

I thank José Silva for helpful discussion and Lars Grotewold for comments on the manuscript and critical references.

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Authors and Affiliations

  • Tilo Kunath
    • 1
  1. 1.Institute for Stem Cell ResearchUniversity of EdinburghEdinburghUK

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