ENU Mutagenesis in the Mouse

Rolf Stottmann1, David Beier2

1 Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, 2 Center for Developmental Biology and Regenerative Medicine, Seattle Children's Research Institute, Seattle, Washington
Publication Name:  Current Protocols in Human Genetics
Unit Number:  Unit 15.4
DOI:  10.1002/0471142905.hg1504s82
Online Posting Date:  July, 2014
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Abstract

ABSTRACT

This unit describes the treatment of laboratory mice with the mutagen N‐ethyl‐N‐nitrosourea (ENU) to induce very highly increased rates of mutation throughout the genome. Further, it describes several popular mating schemes designed to produce animals displaying phenotypes associated with the induced mutations. Curr. Protoc. Hum. Genet. 82:15.4.1‐15.4.10 © 2014 by John Wiley & Sons, Inc.

Keywords: alkylating agent; spermatogonial stem cells; per locus mutation frequency; phenotype‐driven mutagenesis; genome sequencing

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1:

  Materials
  • 95% nondenatured ethanol
  • N‐ethyl‐N‐nitrosourea (ENU; available from Sigma in 1‐g amounts sealed in a 100‐ml serum bottle; an IsoPac also comes with the bottle for temperature control; store at <0°C until use)
  • Phosphate/citrate buffer (see recipe)
  • 8‐ to 10‐week‐old male mice
  • Inactivating solution (see recipe)
  • 1‐, 10‐, and 60‐ml syringes with Luer locks
  • 16‐ and 27‐G needles
  • Disposable cuvettes for spectrophotometric analysis
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Figures

Videos

Literature Cited

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Key References
   Fairfield et al., 2011. See above.
  Good evidence for the ability of exome sequencing to identify ENU mutations.
   Justice et al., 2000. See above.
  The most complete collection of data pertaining to the sensitivity of mouse strains to ENU treatment.
   Weber, J.S. , Salinger, A. , and Justice, M.J. 2000. Optimal N‐ethyl‐N‐nitrosourea (ENU) doses for inbred strains. Genesis 26:230‐233.
  Further useful information about the dose‐response characteristics of mouse inbred lines to ENU treatment.
   Vivian et al., 2002. See above.
  A recent example of how ENU mutagenesis can be applied to embryonic stem cells in culture.
Internet Resources
   http://www.har.mrc.ac.uk
  Mammalian Genetics Unit, Harwell, U.K. ENU Mutagensis Web site.
   http://www.informatics.jax.org/expression.shtml
  Mouse Gene Expression Database (GXD) hosted by the Mouse Genome Informatics program at The Jackson Laboratory.
   http://www.brain‐map.org
  The Allen Brain Atlas Project.
   http://www.gudmap.org
  The GenitoUrinary Molecular Anatomy Project.
   http://www.informatics.jax.org/javawi2/servlet/WIFetch?page=snpQF
  Mouse SNP Query Database from MGI.
   http://www.har.mrc.ac.uk/services/enu‐dna‐archive
  Harwell Mouse ENU DNA Archive.
   http://csbio.unc.edu/CCstatus/Media/MegaMUGAFlyer.pdf
  A recently developed high density SNP panel.
   http://www.sanger.ac.uk/resources/mouse/
  Sanger Institute Mouse Genomics—deep sequencing of multiple inbred mouse strains.
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