Drug-tunable multidimensional synthetic gene control using inducible degron-tagged dCas9 effectors

Dirk A. Kleinjan, Caroline Wardrope, Si Nga Sou and Susan J. Rosser ()
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Dirk A. Kleinjan: Biochemistry, and Biotechnology, SynthSys, School of Biological Sciences, University of Edinburgh
Caroline Wardrope: Biochemistry, and Biotechnology, SynthSys, School of Biological Sciences, University of Edinburgh
Si Nga Sou: Biochemistry, and Biotechnology, SynthSys, School of Biological Sciences, University of Edinburgh
Susan J. Rosser: Biochemistry, and Biotechnology, SynthSys, School of Biological Sciences, University of Edinburgh

Nature Communications, 2017, vol. 8, issue 1, 1-9

Abstract: Abstract The nuclease-deactivated variant of CRISPR-Cas9 proteins (dCas9) fused to heterologous transactivation domains can act as a potent guide RNA sequence-directed inducer or repressor of gene expression in mammalian cells. In such a system the long-term presence of a stable dCas9 effector can be a draw-back precluding the ability to switch rapidly between repressed and activated target gene expression states, imposing a static environment on the synthetic regulatory circuits in the cell. To address this issue we have generated a toolkit of conditionally degradable or stabilisable orthologous dCas9 or Cpf1 effector proteins, thus opening options for multidimensional control of functional activities through combinations of orthogonal, drug-tunable artificial transcription factors.

Date: 2017
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DOI: 10.1038/s41467-017-01222-y

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