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.

  • Brief Communication
  • Published:

Transient cold shock enhances zinc-finger nuclease–mediated gene disruption

Abstract

Zinc-finger nucleases (ZFNs) are powerful tools for editing the genomes of cell lines and model organisms. Given the breadth of their potential application, simple methods that increase ZFN activity, thus ensuring genome modification, are highly attractive. Here we show that transient hypothermia generally and robustly increased the level of stable, ZFN-induced gene disruption, thereby providing a simple technique to enhance the experimental efficacy of ZFNs.

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

Figure 1: Cold shock enhances ZFN activity in human cells.
Figure 2: Effect of the cold shock on ZFNs targeting six genes.

Similar content being viewed by others

References

  1. Morton, J., Davis, M.W., Jorgensen, E.M. & Carroll, D. Proc. Natl. Acad. Sci. USA 103, 16370–16375 (2006).

    Article  CAS  Google Scholar 

  2. Beumer, K., Bhattacharyya, G., Bibikova, M., Trautman, J.K. & Carroll, D. Genetics 172, 2391–2403 (2006).

    Article  CAS  Google Scholar 

  3. Maeder, M.L. et al. Mol. Cell 31, 294–301 (2008).

    Article  CAS  Google Scholar 

  4. Perez, E.E. et al. Nat. Biotechnol. 26, 808–816 (2008).

    Article  CAS  Google Scholar 

  5. Santiago, Y. et al. Proc. Natl. Acad. Sci. USA 105, 5809–5814 (2008).

    Article  CAS  Google Scholar 

  6. Doyon, Y. et al. Nat. Biotechnol. 26, 702–708 (2008).

    Article  CAS  Google Scholar 

  7. Foley, J.E. et al. PLoS One 4, e4348 (2009).

    Article  Google Scholar 

  8. Meng, X., Noyes, M.B., Zhu, L.J., Lawson, N.D. & Wolfe, S.A. Nat. Biotechnol. 26, 695–701 (2008).

    Article  CAS  Google Scholar 

  9. Geurts, A.M. et al. Science 325, 433 (2009).

    Article  CAS  Google Scholar 

  10. Mashimo, T. et al. PLoS One 5, e8870 (2010).

    Article  Google Scholar 

  11. Handel, E.M., Alwin, S. & Cathomen, T. Mol. Ther. 17, 104–111 (2009).

    Article  Google Scholar 

  12. Miller, J.C. et al. Nat. Biotechnol. 25, 778–785 (2007).

    Article  CAS  Google Scholar 

  13. Szczepek, M. et al. Nat. Biotechnol. 25, 786–793 (2007).

    Article  CAS  Google Scholar 

  14. Hockemeyer, D. et al. Nat. Biotechnol. 27, 851–857 (2009).

    Article  CAS  Google Scholar 

  15. Roobol, A., Carden, M.J., Newsam, R.J. & Smales, C.M. FEBS J. 276, 286–302 (2009).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank colleagues at Sangamo Biosciences for testing the cold-shock treatment in their preferred systems and sharing their results, A. Reik, J. Laganière, F. Urnov and E. Rebar for experimental advice, discussion and critical comments on the manuscript and E. Lanphier for support.

Author information

Authors and Affiliations

Authors

Contributions

Y.D., V.M.C., D.F.X. and T.D.V. performed the experiments. Y.D. and M.C.H. designed the experiments. Y.D., M.C.H. and P.D.G. wrote the paper.

Corresponding author

Correspondence to Yannick Doyon.

Ethics declarations

Competing interests

All authors are employees of Sangamo BioSciences.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5 and Supplementary Tables 1–2 (PDF 2938 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Doyon, Y., Choi, V., Xia, D. et al. Transient cold shock enhances zinc-finger nuclease–mediated gene disruption. Nat Methods 7, 459–460 (2010). https://doi.org/10.1038/nmeth.1456

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1456

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