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Related Experiment Videos

Quantifying in vivo somatic mutations using transgenic mouse model systems.

Roy R Swiger1

  • 1Midwest Research Institute, Palm Bay, FL, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 27, 2004
PubMed
Summary

The bacteriophage cII positive selection assay offers a sensitive method for studying mutations in any mouse tissue. This transgenic model system is robust, simple to use, and valuable for mutagenicity assessment.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Toxicology

Background:

  • Transgenic mouse models are crucial for studying in vivo mutagenesis.
  • Existing assays like Big Blue Mouse and MutaMouse have limitations.
  • The bacteriophage cII positive selection assay provides a novel approach.

Purpose of the Study:

  • To describe the application of the bacteriophage cII positive selection assay using the MutaMouse transgenic model.
  • To evaluate its utility for in vivo mutagenesis studies and mutagenicity assessment.

Main Methods:

  • Purification of high-molecular-weight DNA from mouse tissues.
  • In vitro packaging reactions to excise lambda transgenes into viable phage.
  • Adsorption of phage into bacterial hosts for mutation detection via screening or selection assays.

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Main Results:

  • The assay allows for enumeration of mutations in virtually any mouse tissue.
  • Mutant frequencies are determined by the ratio of mutant to total phage units.
  • The method is sensitive, robust, and applicable across various tissues.

Conclusions:

  • The bacteriophage cII positive selection assay in MutaMouse is a sensitive and robust tool for in vivo mutagenesis studies.
  • It facilitates mutagenicity assessment across diverse tissues with accessible laboratory resources.
  • This assay complements existing transgenic models for genotoxicity testing.