Skip to main content

Thank you for visiting 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.

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


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%.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

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


  1. Wiedenheft, B., Sternberg, S.H. & Doudna, J.A. Nature 482, 331–338 (2012).

    CAS  Article  Google Scholar 

  2. Jinek, M. et al. Science 337, 816–821 (2012).

    CAS  Article  Google Scholar 

  3. Cohen, J. Science 332, 784–789 (2011).

    CAS  Article  Google Scholar 

  4. Kim, H., Um, E., Cho, S.R., Jung, C. & Kim, J.S. Nat. Methods 8, 941–943 (2011).

    CAS  Article  Google Scholar 

  5. Kim, H.J., Lee, H.J., Kim, H., Cho, S.W. & Kim, J.S. Genome Res. 19, 1279–1288 (2009).

    CAS  Article  Google Scholar 

  6. Miller, J.C. et al. Nat. Biotechnol. 29, 143–148 (2011).

    CAS  Article  Google Scholar 

  7. Mussolino, C. et al. Nucleic Acids Res. 39, 9283–9293 (2011).

    CAS  Article  Google Scholar 

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

    CAS  Article  Google Scholar 

  9. Kim, E. et al. Genome Res. 22, 1327–1333 (2012).

    CAS  Article  Google Scholar 

  10. Lee, H.J., Kweon, J., Kim, E., Kim, S. & Kim, J.S. Genome Res. 22, 539–548 (2012).

    CAS  Article  Google Scholar 

  11. Lee, H.J., Kim, E. & Kim, J.S. Genome Res. 20, 81–89 (2010).

    CAS  Article  Google Scholar 

  12. Kim, J.S., Lee, H.J. & Carroll, D. Nat. Methods 7, 91, author reply 91–92 (2010).

    CAS  Article  Google Scholar 

  13. Reyon, D. et al. Nat. Biotechnol. 30, 460–465 (2012).

    CAS  Article  Google Scholar 

  14. Cong, L. et al. Science doi:10.1126/science.1231143 (3 January 2013).

  15. Mali, P. et al. Science doi:10.1126/science.1232033 (3 January 2013).

  16. Jiang, W., Bikard, D., Cox, D., Zhang, F. & Marraffini, L.A. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat. Biotechnol. advance online publication, doi:10.1038/nbt.2508 (29 January 2013).

  17. Hwang, W.Y. et al. Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat. Biotechnol. advance online publication, doi:10.1038/nbt.2501 (29 January 2013).

Download references


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

Author information

Authors and Affiliations



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.

Ethics declarations

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)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Further reading


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