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Surrogate reporters for enrichment of cells with nuclease-induced mutations

Abstract

Zinc-finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are powerful tools for creating genetic modifications in eukaryotic cells and organisms. But wild-type and mutant cells that contain genetic modifications induced by these programmable nucleases are often phenotypically indistinguishable, hampering isolation of mutant cells. Here we show that transiently transfected episomal reporters encoding fluorescent proteins can be used as surrogate genes for the efficient enrichment of endogenous gene-modified cells by flow cytometry.

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Figure 1: Principle of the use of surrogate reporters for enrichment of nuclease-modified cells.
Figure 2: A surrogate reporter enriches for TP53-disrupted cells.

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Acknowledgements

J.-S.K. and Hyongbum K. are supported in part by the National Research Foundation of Korea (J.-S.K., 2011-0000402; Hyongbum K., 2011-0013568, 2011-27177, 2011-0019357 and 2011-0028267).

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Authors

Contributions

Hyojin K., E.U., S.-R.C. and C.J. performed the experiments. Hyongbum K. and J.-S.K. wrote the paper.

Corresponding authors

Correspondence to Hyongbum Kim or Jin-Soo Kim.

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Competing interests

J.-S.K. filed a patent application (US 61/445,346) based on this work.

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Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Notes 1–2 (PDF 1479 kb)

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Kim, H., Um, E., Cho, SR. et al. Surrogate reporters for enrichment of cells with nuclease-induced mutations. Nat Methods 8, 941–943 (2011). https://doi.org/10.1038/nmeth.1733

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