Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Reverse genetics by chemical mutagenesis in Caenorhabditis elegans

Abstract

Traditional reverse genetics on yeast, mice and other organisms uses homologous recombination with transgenic DNA to interrupt a target gene. Here we report that target-selected gene inactivation can be achieved in Caenorhabditis elegans with the use of chemical mutagens. We use PCR to selectively visualize deletions in genes of interest; the method is sensitive enough to permit detection of a single mutant among more than 15,000 wild types. A permanent frozen mutant collection of more than a million mutagenized animals has been established, and deletion mutants of several G-protein genes were isolated from it. The approach is suitable to be scaled up for systematic inactivation of all 17,000 C. elegans genes. Because it requires no transgenesis or cell culturing, it may also be applicable to small organisms usually considered to be outside the realm of reverse genetics (for example, other nematodes and insects). Any sequenced gene in any organism that can be handled in very large numbers can possibly be targeted in this way.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Smith, V., Botstein, D. & Brown, P.O. Genetic footprinting: a genomic strategy for determining a gene's function given its sequence. Proc. Natl. Acad. Sci. USA 92, 6479–6483 (1995).

    Article  CAS  Google Scholar 

  2. Smith, V., Chou, K.N., Lashkari, D., Botstein, D. & Brown, P.O. Functional analysis of the genes of yeast chromosome V by genetic footprinting. Science 274, 2069–2074 (1996).

    Article  CAS  Google Scholar 

  3. Waterston, R. & Sulston, J. The genome of Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 92, 10836–10840 (1995).

    Article  CAS  Google Scholar 

  4. Zwaal, R.R., Broeks, A., van Meurs, J., Groenen, J.T.M. & Plasterk, R.H.A. Target-selected gene inactivation in Caenorhabditis elegans by using a frozen transposon insertion mutant bank. Proc. Natl. Acad. Sci. USA 90, 7431–7435 (1993).

    Article  CAS  Google Scholar 

  5. van Luenen, H.G.A.M. & Plasterk, R.H.A. Target site choice of the related transposable elements Tc1 and Tc3 of Caenorhabditis elegans. Nucleic Acids Res. 22, 262–269 (1994).

    Article  CAS  Google Scholar 

  6. Anderson, P., in Caenorhabditis Elegans: Modern Biological Analysis of an Organism (eds Epstein, H.F. & Shakes, D.C.) Methods Cell Biol. 48, 31–58 (1995).

    Book  Google Scholar 

  7. Yandell, M.D., Edgar, L.G. & Wood, W.B. Trimethylpsoralen induces small deletion mutations in Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 91, 1381–1385 (1994).

    Article  CAS  Google Scholar 

  8. Plasterk, R.H.A. Reverse genetics: from gene sequence to mutant worm, in Caenorhabditis elegans: Modern Biological Analysis of an Organism (eds Epstein, H. F. & Shakes, D.C.). Methods Cell Biol. 48, 59–80 (1995).

    Chapter  Google Scholar 

  9. Sulston, J. & Hodgkin, J. Methods. in The Nematode Caenorhabditis Elegans (ed Wood, W. B.) 587–606 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY,) (1988).

    Google Scholar 

  10. Sambrook, J., Fritsch, E.F. & Maniatis, T. Molecular Cloning: A Laboratory Manual, 2nd ed.(Cold Spring Harbor Laboratory, Cold Spring Harbor, New York,) (1989).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ronald H.A. Plasterk.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jansen, G., Hazendonk, E., Thijssen, K. et al. Reverse genetics by chemical mutagenesis in Caenorhabditis elegans. Nat Genet 17, 119–121 (1997). https://doi.org/10.1038/ng0997-119

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng0997-119

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing