Systematic profiling of conditional degron tag technologies for target validation studies.
Systematic profiling of conditional degron tag technologies for target validation studies.
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DOI：
10.1038/s41467-022-33246-4
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发表时间：
2022-09-20
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
条件降解决定子标签 (CDT) 是一种强大的靶标验证工具，它将药理制剂的动力学和可逆作用与遗传操作的普遍性结合起来。然而，CDT 融合蛋白的成功设计通常需要漫长的、临时的构建体设计、失败和重新设计周期。为了解决这个限制，我们在这里报告了一个系统，可以快速比较五个独特 CDT 的活动：AID/AID2、IKZF3d、dTAG、HaloTag 和 SMASh。我们展示了该系统针对 16 种独特蛋白质靶点的实用性。我们发现表达和降解高度依赖于特定的 CDT、构建体设计和靶标。没有一个 CDT 能导致所有靶标的有效表达和/或降解；然而，我们的系统方法能够为每个目标识别至少一个最佳 CDT 融合。为了更广泛地采用 CDT 策略，我们将这些试剂和详细方案作为社区资源提供。条件 Degron 标签是验证和研究新治疗靶点的宝贵工具。在这里，作者比较了 16 种独特蛋白质的 5 个正交标签，并提供了一组载体，供用户系统地筛选具有自己感兴趣的蛋白质的标签。
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 approach enables the identification of at least one optimal CDT fusion for each target. To enable the adoption of CDT strategies more broadly, we have made these reagents, and a detailed protocol, available as a community resource. Conditional Degron Tags are a valuable tool to validate and study novel therapeutic targets. Here, the authors compared 5 orthogonal tags across 16 unique proteins and provide a panel of vectors for users to systematically screen the tags with their own protein of interest.
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影响因子：
14.8
作者：
Arrowsmith CH;Audia JE;Austin C;Baell J;Bennett J;Blagg J;Bountra C;Brennan PE;Brown PJ;Bunnage ME;Buser-Doepner C;Campbell RM;Carter AJ;Cohen P;Copeland RA;Cravatt B;Dahlin JL;Dhanak D;Edwards AM;Frederiksen M;Frye SV;Gray N;Grimshaw CE;Hepworth D;Howe T;Huber KV;Jin J;Knapp S;Kotz JD;Kruger RG;Lowe D;Mader MM;Marsden B;Mueller-Fahrnow A;Müller S;O'Hagan RC;Overington JP;Owen DR;Rosenberg SH;Roth B;Ross R;Schapira M;Schreiber SL;Shoichet B;Sundström M;Superti-Furga G;Taunton J;Toledo-Sherman L;Walpole C;Walters MA;Willson TM;Workman P;Young RN;Zuercher WJ
通讯作者：
Zuercher WJ
影响因子：
64.8
作者：
Erb MA;Scott TG;Li BE;Xie H;Paulk J;Seo HS;Souza A;Roberts JM;Dastjerdi S;Buckley DL;Sanjana NE;Shalem O;Nabet B;Zeid R;Offei-Addo NK;Dhe-Paganon S;Zhang F;Orkin SH;Winter GE;Bradner JE
通讯作者：
Bradner JE
影响因子：
64.5
作者：
McDonald, E. Robert, III;de Weck, Antoine;Sellers, William R.
通讯作者：
Sellers, William R.
影响因子：
4
作者：
Buckley DL;Raina K;Darricarrere N;Hines J;Gustafson JL;Smith IE;Miah AH;Harling JD;Crews CM
通讯作者：
Crews CM
影响因子：
14.8
作者：
Nabet B;Roberts JM;Buckley DL;Paulk J;Dastjerdi S;Yang A;Leggett AL;Erb MA;Lawlor MA;Souza A;Scott TG;Vittori S;Perry JA;Qi J;Winter GE;Wong KK;Gray NS;Bradner JE
通讯作者：
Bradner JE