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CRISPR RNA–guided activation of endogenous human genes

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

References

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

    CAS  Article  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).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  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).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Article  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).

    CAS  Article  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.

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Authors and Affiliations

Authors

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.

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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.

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

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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

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