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CRISPR RNA–guided activation of endogenous human genes

Nature Methods volume 10, pages 977979 (2013) | Download Citation

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.

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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

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

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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.

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.

Corresponding author

Correspondence to J Keith Joung.

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DOI

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

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