Nothing Special   »   [go: up one dir, main page]
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.

  • Brief Communication
  • Published:

CRISPR RNA–guided activation of endogenous human genes

Nature Methods volume 10, pages 977–979 (2013)Cite this article

Subjects

Abstract

Short guide RNAs (gRNAs) can direct catalytically inactive CRISPR-associated 9 nuclease (dCas9) to repress endogenous genes in bacteria and human cells. Here we show that single or multiple gRNAs can direct dCas9 fused to a VP64 transcriptional activation domain to increase expression of endogenous human genes. This proof-of-principle work shows that clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems can target heterologous effector domains to endogenous sites in human cells.

Access through your institution
Buy or subscribe

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

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: RNA-guided activation of endogenous human genes.
Figure 2: Synergistic activation of endogenous human genes by RNA-guided activators.

Similar content being viewed by others

References

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

    Article  CAS  PubMed  Google Scholar 

  2. Cong, L. et al. Science 339, 819–823 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Hwang, W.Y. et al. Nat. Biotechnol. 31, 227–229 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Mali, P. et al. Science 339, 823–826 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Shen, B. et al. Cell Res. 23, 720–723 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Ding, Q. et al. Cell Stem Cell 12, 393–394 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Qi, L.S. et al. Cell 152, 1173–1183 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Bikard, D. et al. Nucleic Acids Res. (doi:10.1093/nar/gkt520) (12 June 2013).

  10. Gilbert, L.A. et al. Cell (doi:10.1016/j.cell.2013.06.044) (9 July 2013).

  11. Beerli, R.R., Segal, D.J., Dreier, B. & Barbas, C.F. Proc. Natl. Acad. Sci. USA 95, 14628–14633 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Maeder, M.L. et al. Nat. Methods 10, 243–245 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Perez-Pinera, P. et al. Nat. Methods 10, 239–242 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Liu, P.-Q. et al. J. Biol. Chem. 276, 11323–11334 (2001).

    Article  CAS  PubMed  Google Scholar 

  15. Fu, Y. et al. Nat. Biotechnol. (doi:10.1038/nbt.2623) (23 June 2013).

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by a US National Institutes of Health (NIH) Director's Pioneer Award DP1 GM105378, NIH R01 NS073124, NIH P50 HG005550, Defense Advanced Research Projects Agency (DARPA) W911NF-11-2-0056, and the Jim and Ann Orr Massachusetts General Hospital Research Scholar Award. We thank J.D. Sander (Massachusetts General Hospital) for providing plasmid pJDS246 and for helpful discussions.

Author information

Authors and Affiliations

  1. Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Morgan L Maeder, Samantha J Linder, Vincent M Cascio, Yanfang Fu, Quan H Ho & J Keith Joung

  2. Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Morgan L Maeder, Samantha J Linder, Vincent M Cascio, Yanfang Fu & J Keith Joung

  3. Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Morgan L Maeder, Samantha J Linder, Vincent M Cascio, Yanfang Fu, Quan H Ho & J Keith Joung

  4. Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, Massachusetts, USA

    Morgan L Maeder & J Keith Joung

  5. Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA

    Yanfang Fu & J Keith Joung

Authors
  1. Morgan L Maeder
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Samantha J Linder
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vincent M Cascio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yanfang Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Quan H Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J Keith Joung
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.L.M. and J.K.J. conceived of the study, designed the experiments and wrote the manuscript. M.L.M., S.J.L., V.M.C., Y.F. and Q.H.H. performed experiments.

Corresponding author

Correspondence to J Keith Joung.

Ethics declarations

Competing interests

M.L.M. and J.K.J. are inventors on patent applications (61/838,148 and 61/799,647) describing the dCas9-VP64 fusion protein and its use to activate gene expression. J.K.J. has a financial interest in Transposagen Biopharmaceuticals. J.K.J.'s interests were reviewed and are managed by Massachusetts General Hospital and Partners HealthCare in accordance with their conflict of interest policies.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, and Supplementary Tables 1 and 2 (PDF 290 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Maeder, M., Linder, S., Cascio, V. et al. CRISPR RNA–guided activation of endogenous human genes. Nat Methods 10, 977–979 (2013). https://doi.org/10.1038/nmeth.2598

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.2598

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