Skip to main content

Thank you for visiting nature.com. 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.

A split-Cas9 architecture for inducible genome editing and transcription modulation

Nature Biotechnology volume 33, pages 139–142 (2015)Cite this article

Subjects

To the Editor:

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

Buy

All prices are NET prices.

Figure 1: Generation and optimization of inducible split-Cas9 fragments.
Figure 2: Inducible transcriptional activation using split dCas9-VP64 fusions.

References

  1. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013).

    Article  CAS  Google Scholar 

  2. Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013).

    Article  CAS  Google Scholar 

  3. Konermann, S. et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 10.1038/nature14136 (10 December 2014).

  4. Gilbert, L.A. et al. Genome-scale CRISPR-mediated control of gene repression and activation. Cell 159, 647–661 (2014).

    Article  CAS  Google Scholar 

  5. Qi, L.S. et al. Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell 152, 1173–1183 (2013).

    Article  CAS  Google Scholar 

  6. Nishimasu, H. et al. Crystal structure of cas9 in complex with guide RNA and target DNA. Cell 156, 935–949 (2014).

    Article  CAS  Google Scholar 

  7. Banaszynski, L.A., Liu, C.W. & Wandless, T.J. Characterization of the FKBP·rapamycin·FRB ternary complex. J. Am. Chem. Soc. 127, 4715–4721 (2005).

    Article  CAS  Google Scholar 

  8. Konermann, S. et al. Optical control of mammalian endogenous transcription and epigenetic states. Nature 500, 472–476 (2013).

    Article  CAS  Google Scholar 

  9. Hsu, P.D. et al. DNA targeting specificity of RNA-guided Cas9 nucleases. Nat. Biotechnol. 31, 827–832 (2013).

    Article  CAS  Google Scholar 

  10. Fu, Y., Sander, J.D., Reyon, D., Cascio, V.M. & Joung, J.K. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat. Biotechnol. 32, 279–284 (2014).

    Article  CAS  Google Scholar 

  11. Perez-Pinera, P. et al. RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Nat. Methods 10, 973–976 (2013).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank N.E. Sanjana, S. Konermann, M.D. Brigham, A. Trevino, F.A. Ran and W. Yan for technical assistance, S. Jones, M. Heidenreich and K. Zheng for editing and members of the Zhang laboratory for discussion, support and advice. F.Z. is supported by the National Institute of Mental Health (NIMH) through a National Institutes of Health (NIH) Director's Pioneer Award (DP1-MH100706), the National Institute of Neurological Disorders and Stroke (NINDS) through an NIH Transformative R01 grant (R01-NS 07312401), National Science Foundation (NSF) Waterman Award, the Keck, Damon Runyon, Searle Scholars, Klingenstein, Vallee, Merkin and Simons Foundations, and Bob Metcalfe, New York Stem Cell Foundation. F.Z. is a New York Stem Cell Foundation Robertson Investigator. CRISPR reagents are available to the academic community through Addgene, and associated protocols, support forum and computational tools are available via the Zhang laboratory website (http://www.genome-engineering.org).

Author information

Authors and Affiliations

  1. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Bernd Zetsche, Sara E Volz & Feng Zhang

  2. McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Bernd Zetsche, Sara E Volz & Feng Zhang

  3. Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Bernd Zetsche, Sara E Volz & Feng Zhang

  4. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Bernd Zetsche, Sara E Volz & Feng Zhang

  5. Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Bonn, Germany

    Bernd Zetsche

Authors
  1. Bernd Zetsche
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sara E Volz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Feng Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

B.Z. and F.Z. conceived the project and designed the experiments. B.Z. and S.E.V. performed experiments and analyzed data. B.Z. and F.Z. wrote the paper with help from all authors.

Corresponding author

Correspondence to Feng Zhang.

Ethics declarations

Competing interests

Feng Zhang is a scientific advisor of Editas Medicine and of Horizon Discovery.

Supplementary information

Supplementary Figures and Text

Supplementary Figures 1 and 2, Supplementary Tables 1–5, Supplementary Methods (PDF 1641 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zetsche, B., Volz, S. & Zhang, F. A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol 33, 139–142 (2015). https://doi.org/10.1038/nbt.3149

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt.3149

This article is cited by

Search

Advanced search

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