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Activation of stably silenced genes by recruitment of a synthetic de-methylating module

Author

Listed:
  • Wing Fuk Chan

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne
    Monash University)

  • Hannah D. Coughlan

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne)

  • Yunshun Chen

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne)

  • Christine R. Keenan

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne)

  • Gordon K. Smyth

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne
    The University of Melbourne)

  • Andrew C. Perkins

    (Monash University)

  • Timothy M. Johanson

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne)

  • Rhys S. Allan

    (The Walter and Eliza Hall Institute of Medical Research
    The University of Melbourne)

Abstract

Stably silenced genes that display a high level of CpG dinucleotide methylation are refractory to the current generation of dCas9-based activation systems. To counter this, we create an improved activation system by coupling the catalytic domain of DNA demethylating enzyme TET1 with transcriptional activators (TETact). We show that TETact demethylation-coupled activation is able to induce transcription of suppressed genes, both individually and simultaneously in cells, and has utility across a number of cell types. Furthermore, we show that TETact can effectively reactivate embryonic haemoglobin genes in non-erythroid cells. We anticipate that TETact will expand the existing CRISPR toolbox and be valuable for functional studies, genetic screens and potential therapeutics.

Suggested Citation

  • Wing Fuk Chan & Hannah D. Coughlan & Yunshun Chen & Christine R. Keenan & Gordon K. Smyth & Andrew C. Perkins & Timothy M. Johanson & Rhys S. Allan, 2022. "Activation of stably silenced genes by recruitment of a synthetic de-methylating module," Nature Communications, Nature, vol. 13(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33181-4
    DOI: 10.1038/s41467-022-33181-4
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    References listed on IDEAS

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    1. Silvana Konermann & Mark D. Brigham & Alexandro E. Trevino & Julia Joung & Omar O. Abudayyeh & Clea Barcena & Patrick D. Hsu & Naomi Habib & Jonathan S. Gootenberg & Hiroshi Nishimasu & Osamu Nureki &, 2015. "Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex," Nature, Nature, vol. 517(7536), pages 583-588, January.
    2. Daniel M. Sapozhnikov & Moshe Szyf, 2021. "Unraveling the functional role of DNA demethylation at specific promoters by targeted steric blockage of DNA methyltransferase with CRISPR/dCas9," Nature Communications, Nature, vol. 12(1), pages 1-26, December.
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