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

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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.

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


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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

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.

Correspondence to Yannick Doyon.

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All authors are employees of Sangamo BioSciences.

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

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