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Lookup NU author(s): Dr Chiara ManiaciORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2022, Crown. Modulation of protein abundance using tag-Targeted Protein Degrader (tTPD) systems targeting FKBP12F36V (dTAGs) or HaloTag7 (HaloPROTACs) are powerful approaches for preclinical target validation. Interchanging tags and tag-targeting degraders is important to achieve efficient substrate degradation, yet limited degrader/tag pairs are available and side-by-side comparisons have not been performed. To expand the tTPD repertoire we developed catalytic NanoLuc-targeting PROTACs (NanoTACs) to hijack the CRL4CRBN complex and degrade NanoLuc tagged substrates, enabling rapid luminescence-based degradation screening. To benchmark NanoTACs against existing tTPD systems we use an interchangeable reporter system to comparatively test optimal degrader/tag pairs. Overall, we find the dTAG system exhibits superior degradation. To align tag-induced degradation with physiology we demonstrate that NanoTACs limit MLKL-driven necroptosis. In this work we extend the tTPD platform to include NanoTACs adding flexibility to tTPD studies, and benchmark each tTPD system to highlight the importance of comparing each system against each substrate.
Author(s): Grohmann C, Magtoto CM, Walker JR, Chua NK, Gabrielyan A, Hall M, Cobbold SA, Mieruszynski S, Brzozowski M, Simpson DS, Dong H, Dorizzi B, Jacobsen AV, Morrish E, Silke N, Murphy JM, Heath JK, Testa A, Maniaci C, Ciulli A, Lessene G, Silke J, Feltham R
Publication type: Article
Publication status: Published
Journal: Nature Communications
Year: 2022
Volume: 13
Issue: 1
Online publication date: 19/04/2022
Acceptance date: 25/03/2022
Date deposited: 04/05/2022
ISSN (electronic): 2041-1723
Publisher: Nature Publishing Group
URL: https://doi.org/10.1038/s41467-022-29670-1
DOI: 10.1038/s41467-022-29670-1
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