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Synergistic and tunable human gene activation by combinations of synthetic transcription factors

Nature Methods volume 10, pages 239–242 (2013)Cite this article

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Abstract

Mammalian genes are regulated by the cooperative and synergistic actions of many transcription factors. In this study we recapitulate this complex regulation in human cells by targeting endogenous gene promoters, including regions of closed chromatin upstream of silenced genes, with combinations of engineered transcription activator–like effectors (TALEs). These combinations of TALE transcription factors induced substantial gene activation and allowed tuning of gene expression levels that will broadly enable synthetic biology, gene therapy and biotechnology.

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Figure 1: Synergistic activation of gene expression by combinations of TALE-TFs.
Figure 2: Combinatorial regulation of gene expression by TALE-TFs.

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Acknowledgements

This work was supported by a US National Institutes of Health (NIH) Director's New Innovator Award (DP2-OD008586), a National Science Foundation (NSF) Faculty Early Career Development (CAREER) Award (CBET-1151035), NIH R03-AR061042, The Hartwell Foundation Individual Biomedical Research Award and a March of Dimes Basil O'Connor Starter Scholar Award to C.A.G.; grants from the NIH (P50-GM081883) and the Defense Advance Research Projects Agency (HR0011-09-1-0040) to A.J.H.; and grants from the NIH to G.E.C. (U54-HG004563) and F.G. (R01-AR048852). D.G.O. was supported by a predoctoral fellowship from the American Heart Association. K.A.G. was supported by an NSF Graduate Research Fellowship.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA

    Pablo Perez-Pinera, David G Ousterout, Jonathan M Brunger, Katherine A Glass, Farshid Guilak & Charles A Gersbach

  2. Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA

    Jonathan M Brunger, Katherine A Glass & Farshid Guilak

  3. Department of Medicine, Duke University Medical Center, Durham, North Carolina, USA

    Alicia M Farin

  4. Institute for Genome Sciences and Policy, Duke University, Durham, North Carolina, USA

    Gregory E Crawford, Alexander J Hartemink & Charles A Gersbach

  5. Department of Pediatrics, Division of Medical Genetics, Duke University Medical Center, Durham, North Carolina, USA

    Gregory E Crawford

  6. Department of Computer Science, Duke University, Durham, North Carolina, USA

    Alexander J Hartemink

Authors
  1. Pablo Perez-Pinera
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  2. David G Ousterout
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  3. Jonathan M Brunger
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  4. Alicia M Farin
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  5. Katherine A Glass
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  6. Farshid Guilak
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  7. Gregory E Crawford
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  8. Alexander J Hartemink
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  9. Charles A Gersbach
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Contributions

P.P.-P., A.M.F., G.E.C., A.J.H. and C.A.G. designed experiments. P.P.-P., D.G.O., J.M.B., A.M.F. and K.A.G. performed the experiments. P.P.-P., F.G., G.E.C., A.J.H. and C.A.G. analyzed the data. P.P.-P. and C.A.G. wrote the manuscript.

Corresponding author

Correspondence to Charles A Gersbach.

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The authors declare no competing financial interests.

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Supplementary Figures 1–7 and Supplementary Tables 1–3 (PDF 6092 kb)

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Perez-Pinera, P., Ousterout, D., Brunger, J. et al. Synergistic and tunable human gene activation by combinations of synthetic transcription factors. Nat Methods 10, 239–242 (2013). https://doi.org/10.1038/nmeth.2361

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