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..
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11541768 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2024.103413 | DOI Listing |
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