2. Academic Validation
  3. Systematic profiling of conditional degron tag technologies for target validation studies

Systematic profiling of conditional degron tag technologies for target validation studies

  • Nat Commun. 2022 Sep 20;13(1):5495. doi: 10.1038/s41467-022-33246-4.
Daniel P Bondeson 1,  Zachary Mullin-Bernstein 1,  Sydney Oliver 1,  Thomas A Skipper 1,  Thomas C Atack 1,  Nolan Bick 1,  Meilani Ching 1,  Andrew A Guirguis 1 2 3 4,  Jason Kwon 1 2,  Carly Langan 1,  Dylan Millson 1 2,  Brenton R Paolella 1,  Kevin Tran 3 4,  Sarah J Wie 1,  Francisca Vazquez 1,  Zuzana Tothova 1 2,  Todd R Golub 1 2,  William R Sellers # 1 2,  Alessandra Ianari # 5
Affiliations

Affiliations

  • 1 The Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • 2 Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA.
  • 3 Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
  • 4 Sir Peter MacCallum Department of Oncology, The University of Melbourne, Melbourne, VIC, Australia.
  • 5 The Broad Institute of MIT and Harvard, Cambridge, MA, USA. [email protected].
  • # Contributed equally.
PMID: 36127368 DOI: 10.1038/s41467-022-33246-4
Abstract

Conditional degron tags (CDTs) are a powerful tool for target validation that combines the kinetics and reversible action of pharmacological agents with the generalizability of genetic manipulation. However, successful design of a CDT fusion protein often requires a prolonged, ad hoc cycle of construct design, failure, and re-design. To address this limitation, we report here a system to rapidly compare the activity of five unique CDTs: AID/AID2, IKZF3d, dTAG, HaloTag, and SMASh. We demonstrate the utility of this system against 16 unique protein targets. We find that expression and degradation are highly dependent on the specific CDT, the construct design, and the target. None of the CDTs leads to efficient expression and/or degradation across all targets; however, our systematic approach enables the identification of at least one optimal CDT fusion for each target. To enable the adoption of CDT strategies more broadly, we have made these reagents, and a detailed protocol, available as a community resource.

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