The dTAG system for immediate and target-specific protein degradation
By Behnam Nabet, Justin M. Roberts, Dennis L. Buckley, Joshiawa Paulk, S. Dastjerdi, A. Yang, Alan L. Leggett, Michael A. Erb, Matthew A. Lawlor, Amanda L. Souza and 8 more. Nature Chemical Biology, 2018
886 citations recorded by the source corpus.
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.The dTAG system pairs potent heterobifunctional degraders and extensible tagging strategies to achieve immediate and reversible degradation of divergent proteins, facilitating biological investigation and drug target validation in cells and in mice.
Subfields: Molecular Biology, Oncology
View Paper: https://www.semanticscholar.org/paper/017419b08ce5005684058db76af788f08eba956f
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Citations in the corpus
- Proteolysis Targeting Chimeras as Therapeutics and Tools for Biological Discovery (2020, positive): “While PROTACs may need optimization of their physico-chemical properties for in vivo work, there are many examples of PROTACs functioning in vivo, including mammals (Bondeson et al., 2015; Burslem et al., 2018c; Jain et al., 2019; Nabet et al., 2018; Winter et al., 2015) and non-human primates (Sun ”
- Transcription factors activate genes through the phase separation capacity of their activation domains (2018, methodology): “We investigated whether the Mediator condensates present at SEs are dependent on OCT4 using a degradation strategy (Nabet et al., 2018).”
- PROteolysis TArgeting Chimeras (PROTACs) - Past, Present and Future (2019, background): “This has been done in a recent example, there CRISPR-mediated knock-in of FKBP12F36V provided endogenous expressed FKBP12F36V-POI fusion protein [87].”