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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.
Details
; Roberts, Justin M 2
; Buckley, Dennis L 3 ; Paulk, Joshiawa 3 ; Dastjerdi, Shiva 2 ; Annan Yang 2 ; Leggett, Alan L 4 ; Erb, Michael A 2
; Lawlor, Matthew A 2 ; Souza, Amanda 3 ; Scott, Thomas G 2 ; Vittori, Sarah 2 ; Perry, Jennifer A 2 ; Qi, Jun 5 ; Winter, Georg E 6 ; Kwok-Kin Wong 7 ; Gray, Nathanael S 1
; Bradner, James E 8
1 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA
2 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA
3 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA; Novartis Institutes for BioMedical Research, Cambridge, MA, USA
4 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA
5 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Medicine, Harvard Medical School, Boston, MA, USA
6 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA; 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 Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Medicine, Harvard Medical School, Boston, MA, USA; Novartis Institutes for BioMedical Research, Cambridge, MA, USA