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An antibiotic selection marker for nematode transgenesis

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Abstract

We have developed a nematode transformation vector carrying the bacterial neomycin resistance gene (NeoR) and shown that it could confer resistance to G-418 on both wild-type Caenorhabditis elegans and C. briggsae. This selection system allows hands-off maintenance and enrichment of transgenic worms carrying non-integrated transgenes on selective plates. We also show that this marker can be used for Mos1-mediated single-copy insertion in wild-type genetic backgrounds (MosSCI-biotic).

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Figure 1: NeoR as a genetic marker for nematode transgenesis.
Figure 2: Enrichment of NeoR transgenic lines in the presence of G-418.

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Acknowledgements

Supported by the Program INSERM “Avenir” (D.D.), la Fondation Bettencourt-Schueller (D.D.), le Conseil Régional d'Aquitaine (D.D.), la Fondation pour la Recherche Médicale (D.D.), Natural Science and Engineering Research Council of Canada (D.B.) and le Ministère Français de l'Enseignement de la Recherche et des Technologies (R.G.-S.). We thank I.A. Hope, J. Ewbanks and J.L. Bessereau for discussions and access to facilities; and T. Leste-Lasserre and G. Drut for discussion about qPCR. EN5271 and EN5273 and MosSCI related plasmids were provided by J.L. Bessereau (INSERM U1024, Institute of Biology of the École Normale Supérieure). The Caenorhabditis Genetics Center, which is funded by the National Center for Research Resources of the US National Institutes of Health, provided the nematode species.

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Authors and Affiliations

  1. Genome Regulation and Evolution, INSERM U869, Institut Européen de Chimie et Biologie, Pessac, France

    Rosina Giordano-Santini & Denis Dupuy

  2. Center for Cancer Systems Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Stuart Milstein, Nenad Svrzikapa & Marc Vidal

  3. Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Stuart Milstein, Nenad Svrzikapa & Marc Vidal

  4. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Stuart Milstein, Nenad Svrzikapa & Marc Vidal

  5. Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia, Canada

    Domena Tu, Robert Johnsen & David Baillie

Authors
  1. Rosina Giordano-Santini
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  2. Stuart Milstein
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  3. Nenad Svrzikapa
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  4. Domena Tu
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  5. Robert Johnsen
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  6. David Baillie
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  7. Marc Vidal
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  8. Denis Dupuy
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Contributions

S.M. and N.S. performed preliminary experiments under the supervision of M.V. D.D. designed and supervised the project and constructed the pDD04neo vector with N.S. Microinjections were performed by R.G.-S. and D.T. under the supervision of D.D., D.B. and R.J. R.G.-S. constructed MosSCI-biotic related vectors, characterized the transgenic worms and wrote the manuscript with D.D.

Corresponding author

Correspondence to Denis Dupuy.

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The authors declare no competing financial interests.

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Supplementary Text and Figures

Supplementary Figures 1– 3 and Supplementary Tables 1–7 (PDF 738 kb)

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Giordano-Santini, R., Milstein, S., Svrzikapa, N. et al. An antibiotic selection marker for nematode transgenesis. Nat Methods 7, 721–723 (2010). https://doi.org/10.1038/nmeth.1494

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  • DOI: https://doi.org/10.1038/nmeth.1494

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