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Targeted gene deletions in C. elegans using transposon excision

Nature Methods volume 7, pages 451–453 (2010)Cite this article

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Abstract

We developed a method, MosDEL, to generate targeted knockouts of genes in Caenorhabditis elegans by injection. We generated a double-strand break by mobilizing a Mos1 transposon adjacent to the region to be deleted; the double-stranded break is repaired using injected DNA as a template. Repair can delete up to 25 kb of DNA and simultaneously insert a positive selection marker.

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Figure 1: Using Mos1 transposons to create targeted deletions.
Figure 2: Deleting lethal genes and multiple similar genes.

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Change history

  • 06 May 2010

    In the version of this article initially published online, the statement that MosDEL can delete 10,069 of 10,154 genes presently lacking a deletion allele was an underestimate; 15,072 of the 15,157 genes presently lacking a publicly available deletion allele can be deleted by MosDEL. Also, the species origin of the Mos1 transposon was incorrect and the abbreviation for Mos1 excision-induced transgene-instructed gene conversion was incorrect. These errors have been corrected online for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank L. Segalat (Centre National de la Recherche Scientifique, Lyon, France) and members of the NemaGENETAG consortium for Mos1 strains. C.F.-J. is funded by a fellowship from the Lundbeck Foundation and G.H. by a Jane Coffin Childs fellowship funded by Howard Hughes Medical Institute. M.B. is funded by a McKnight Grant. D.G.M. is funded by Genome Canada and Genome British Columbia.

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Authors and Affiliations

  1. Department of Biology, Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah, USA

    Christian Frøkjær-Jensen, M Wayne Davis, Gunther Hollopeter, Rachel Lofgren, Michael Prestgard-Duke & Erik M Jorgensen

  2. Department of Biomedical Sciences and Danish National Research Foundation Centre for Cardiac Arrhythmia, University of Copenhagen, Copenhagen, Denmark

    Christian Frøkjær-Jensen

  3. Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada

    Jon Taylor & Donald G Moerman

  4. Hi-Line Informatics, Livingston, Montana, USA

    Todd W Harris

  5. Department of Biology, University of Utah, Salt Lake City, Utah, USA

    Paola Nix & Michael Bastiani

Authors
  1. Christian Frøkjær-Jensen
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Contributions

C.F.-J., M.W.D., G.H. and E.M.J. conceived and designed experiments; C.F.-J., G.H., J.T., P.N., R.L. and M.P.-D. performed experiments; T.W.H. performed the bioinformatic analysis of Mos1 distribution; M.B., D.G.M. and E.M.J. provided supervision and funding; C.F.-J. and E.M.J. wrote the manuscript.

Corresponding author

Correspondence to Erik M Jorgensen.

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Competing interests

E.M.J. is the author of a patent covering techniques described in this paper (US patent 7,196,244 and European patent pending). T.W.H. is the sole proprietor of Hi-Line Informatics.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–2 and Supplementary Tables 1,3 (PDF 1698 kb)

Supplementary Table 2

Mos1 distribution. (XLS 12467 kb)

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Frøkjær-Jensen, C., Davis, M., Hollopeter, G. et al. Targeted gene deletions in C. elegans using transposon excision. Nat Methods 7, 451–453 (2010). https://doi.org/10.1038/nmeth.1454

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