Protocol to identify E3 ligases amenable to biodegraders using a cell-based screening
Here, we provide a protocol for the identification of E3 ubiquitin ligases that are functional when implemented as biodegraders using a cell-based screening assay. We describe steps for establishing a stable cell line expressing a GFP-tagged protein of interest (POI), preparing a sub-library of E3 ligases to screen, and performing the cell-based screening. This protocol can be broadly applied to identify any functional E3 ligase in a biodegrader setting. For complete details on the use and execution of this protocol, please refer to Cornebois et al.<sup>1</sup>.
Authors
- Klervi Desrumeaux
- Nicolas Béry (ORCID: https://orcid.org/0000-0002-2643-3897)
- Marie Sorbara
- Pierre Cordelier (ORCID: https://orcid.org/0000-0003-2096-7967)
- Anaïs Cornebois
- Margot Cristol
Institutions
- Centre National de la Recherche Scientifique (FR)
- Université Toulouse III - Paul Sabatier (FR)
- Inserm (FR)
- Université Fédérale de Toulouse Midi-Pyrénées (FR)
- Sanofi (France) (FR)
- Centre de Recherches en Cancérologie de Toulouse (FR)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-10-24
- DOI
- https://doi.org/10.1016/j.xpro.2024.103413
- Primary Topic
- Protein Degradation and Inhibitors
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Sanofi
- Institut National de la Santé et de la Recherche Médicale
- Association Nationale de la Recherche et de la Technologie
- Ligue Contre le Cancer
- Fondation de France
- Inserm Transfert