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Efficient generation and mapping of recessive developmental mutations using ENU mutagenesis

Nature Genetics volume 30, pages 185–189 (2002)Cite this article

Abstract

Treatment with N-ethyl-N-nitrosourea (ENU) efficiently generates single-nucleotide mutations in mice1,2,3. Along with the renewed interest in this approach, much attention has been given recently to large screens with broad aims4,5; however, more finely focused studies have proven very productive as well6,7,8. Here we show how mutagenesis together with genetic mapping can facilitate the rapid characterization of recessive loci required for normal embryonic development. We screened third-generation progeny of mutagenized mice at embryonic day (E) 18.5 for abnormalities of organogenesis. We ascertained 15 monogenic mutations in the 54 families that were comprehensively analyzed. We carried out the experiment as an outcross, which facilitated the genetic mapping of the mutations by haplotype analysis. We mapped seven of the mutations and identified the affected locus in two lines. Using a hierarchical approach, it is possible to maximize the efficiency of this analysis so that it can be carried out easily with modest infrastructure and resources.

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Figure 1: Line 156: Ryr1m1Bei.
Figure 2: Line 104: shorty.
Figure 3: Line 172: Pkd1m1Bei.
Figure 4: Line 158: alien.

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Acknowledgements

We thank K. Simms, J. Kim, S. Kuida and A. Salinger for their contributions to this project. This work was supported by grants from the National Institutes of Health (M.J.J. and D.R.B.).

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Authors and Affiliations

  1. Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Bruce J. Herron, Weining Lu, Cherie Rao, Shanming Liu, Heiko Peters, Roderick T. Bronson & David R. Beier

  2. Tufts University School of Veterinary Medicine, North Grafton, Massachusetts, USA

    Roderick T. Bronson

  3. Baylor College of Medicine, Houston, Texas, USA

    Monica J. Justice

  4. Wichita State University, Wichita, Kansas, USA

    J. David McDonald

Authors
  1. Bruce J. Herron
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  2. Weining Lu
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  3. Cherie Rao
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  5. Heiko Peters
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  6. Roderick T. Bronson
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  7. Monica J. Justice
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  8. J. David McDonald
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  9. David R. Beier
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Correspondence to David R. Beier.

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Herron, B., Lu, W., Rao, C. et al. Efficient generation and mapping of recessive developmental mutations using ENU mutagenesis. Nat Genet 30, 185–189 (2002). https://doi.org/10.1038/ng812

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