When the underlying biological functions of a novel protein of interest (POI) are unknown, the consequences of its perturbation are also unpredictable. Many targets arise from genome-wide association studies or large-scale functional experiments 1, 2 and are often poorly characterized, bringing exciting opportunities and concomitant challenges.

To enable the adoption of CDT strategies more broadly, we have made these reagents, and a detailed protocol, available as a community resource.Ī major focus in biomedical research is the discovery of novel therapeutic targets. 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. We find that expression and degradation are highly dependent on the specific CDT, the construct design, and the target. We demonstrate the utility of this system against 16 unique protein targets. 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. However, successful design of a CDT fusion protein often requires a prolonged, ad hoc cycle of construct design, failure, and re-design. 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.

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