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