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Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease

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Abstract

We employ the CRISPR-Cas system of Streptococcus pyogenes as programmable RNA-guided endonucleases (RGENs) to cleave DNA in a targeted manner for genome editing in human cells. We show that complexes of the Cas9 protein and artificial chimeric RNAs efficiently cleave two genomic sites and induce indels with frequencies of up to 33%.

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Figure 1: RGEN-catalyzed cleavage of plasmid DNA in vitro and in cellula.
Figure 2: RGEN-driven mutations at endogenous chromosomal sites.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (2012-0001225) and ToolGen, Inc. We thank Jae Kyung Chon for bioinformatic analysis.

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Author notes

  1. Seung Woo Cho and Sojung Kim: These authors contributed equally to this work.

Authors and Affiliations

  1. National Creative Research Initiatives Center for Genome Engineering, Seoul National University, Seoul, South Korea

    Seung Woo Cho, Sojung Kim, Jong Min Kim & Jin-Soo Kim

  2. Department of Chemistry, Seoul National University, Seoul, South Korea

    Seung Woo Cho, Sojung Kim, Jong Min Kim & Jin-Soo Kim

Authors
  1. Seung Woo Cho
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  2. Sojung Kim
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  3. Jong Min Kim
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  4. Jin-Soo Kim
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Contributions

S.W.C., S.K. and J.M.K. performed the experiments. J.-S.K. wrote the manuscript.

Corresponding author

Correspondence to Jin-Soo Kim.

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Competing interests

S.W.C., S.K. and J.-S.K. have filed a patent based on this work.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Methods and Supplementary Table 1 (PDF 550 kb)

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Cho, S., Kim, S., Kim, J. et al. Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease. Nat Biotechnol 31, 230–232 (2013). https://doi.org/10.1038/nbt.2507

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