biostudies-literatureBondeson DPNCI NIH HHSCenter for Strategic Scientific Initiatives, National Cancer Institute5495https://www.ebi.ac.uk/biostudies/studies/S-EPMC9489723biostudies-literatureUnknown13(1)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 approachNature communicationsSystematic profiling of conditional degron tag technologies for target validation studies.PMC9489723R01 CA233626R35 CA242457CA212229K00 CA212229CA233626CA242457Mullin-Bernstein ZSellers WRKwon JGuirguis AABick NSkipper TAWie SJBondeson DPAtack TCPaolella BRTothova ZIanari AOliver SVazquez FLangan CTran KGolub TRChing MMillson DfalseSystematic profiling of conditional degron tag technologies for target validation studies.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 approach2022-01-01T00:00:00Z2022 Sep2026-05-31T11:03:35.194Z2024-11-20T15:22:16.667ZS-EPMC94897233612736810.1038/s41467-022-33246-4