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Surrogate reporters for enrichment of cells with nuclease-induced mutations

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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 and Affiliations

  1. Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea

    Hyojin Kim & Jin-Soo Kim

  2. Cha Stem Cell Institute, Cha University, Seoul, South Korea

    Eunji Um

  3. Department and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, Seoul, South Korea

    Sung-Rae Cho

  4. Graduate School of Biomedical Science and Engineering/College of Medicine, Hanyang University, Sungdong-gu, Seoul, South Korea.,

    Chorong Jung & Hyongbum Kim

Authors
  1. Hyojin Kim
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  2. Eunji Um
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  3. Sung-Rae Cho
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  4. Chorong Jung
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  5. Hyongbum Kim
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  6. Jin-Soo Kim
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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 Text and Figures

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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  • DOI: https://doi.org/10.1038/nmeth.1733

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