2. Academic Validation
  3. The dTAG system for immediate and target-specific protein degradation

The dTAG system for immediate and target-specific protein degradation

  • Nat Chem Biol. 2018 May;14(5):431-441. doi: 10.1038/s41589-018-0021-8.
Behnam Nabet 1 2,  Justin M Roberts 3,  Dennis L Buckley 3 4,  Joshiawa Paulk 3 4,  Shiva Dastjerdi 3,  Annan Yang 3,  Alan L Leggett 1,  Michael A Erb 3,  Matthew A Lawlor 3,  Amanda Souza 3 4,  Thomas G Scott 3,  Sarah Vittori 3,  Jennifer A Perry 3,  Jun Qi 1 5,  Georg E Winter 3 6,  Kwok-Kin Wong 7,  Nathanael S Gray 8 9,  James E Bradner 10 11 12
Affiliations

Affiliations

  • 1 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA.
  • 2 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
  • 3 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
  • 4 Novartis Institutes for BioMedical Research, Cambridge, MA, USA.
  • 5 Department of Medicine, Harvard Medical School, Boston, MA, USA.
  • 6 CeMM- Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • 7 Laura and Isaac Perlmutter Cancer Center, NYU Langone Medical Center, New York, NY, USA.
  • 8 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA. nathanael_gray@dfci.harvard.edu.
  • 9 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. nathanael_gray@dfci.harvard.edu.
  • 10 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA. james.bradner@novartis.com.
  • 11 Department of Medicine, Harvard Medical School, Boston, MA, USA. james.bradner@novartis.com.
  • 12 Novartis Institutes for BioMedical Research, Cambridge, MA, USA. james.bradner@novartis.com.
PMID: 29581585 DOI: 10.1038/s41589-018-0021-8
Abstract

Dissection of complex biological systems requires target-specific control of the function or abundance of proteins. Genetic perturbations are limited by off-target effects, multicomponent complexity, and irreversibility. Most limiting is the requisite delay between modulation to experimental measurement. To enable the immediate and selective control of single protein abundance, we created a chemical biology system that leverages the potency of cell-permeable heterobifunctional degraders. The dTAG system pairs a novel degrader of FKBP12F36V with expression of FKBP12F36V in-frame with a protein of interest. By transgene expression or CRISPR-mediated locus-specific knock-in, we exemplify a generalizable strategy to study the immediate consequence of protein loss. Using dTAG, we observe an unexpected superior antiproliferative effect of pan-BET bromodomain degradation over selective BRD4 degradation, characterize immediate effects of KRASG12V loss on proteomic signaling, and demonstrate rapid degradation in vivo. This technology platform will confer kinetic resolution to biological investigation and provide target validation in the context of drug discovery.

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