The dTAG system for immediate and target-specific protein degradation.
The dTAG system for immediate and target-specific protein degradation.
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DOI：
10.1038/s41589-018-0021-8
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发表时间：
2018-05
影响因子：
14.8
通讯作者：
Bradner JE
中科院分区：
文献类型：
--
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
剖析复杂的生物系统需要对蛋白质功能或丰度进行目标特异性控制。遗传扰动受到脱靶效应、多组分复杂性和不可逆性的限制。最大的限制是从调制到实验测量的必要延迟。为了能够立即和选择性地控制单个蛋白质丰度，我们创建了一个化学生物学系统，该系统利用了细胞可渗透的异双功能降解剂的效力。dTAG系统将FKBP12F36V的新型降解剂与FKBP12F36V与感兴趣蛋白的框内表达配对。通过转基因表达或CRISPR介导的基因座特异性敲入，我们提出了一种可推广的策略来研究蛋白质丢失的直接后果。使用dTAG，我们观察到泛BET布罗莫结构域降解相对于选择性BRD 4降解的出乎意料的上级抗增殖作用，表征KRASG 12 V损失对蛋白质组信号传导的直接作用，并证明体内快速降解。该技术平台将为生物学研究提供动力学分辨率，并在药物发现的背景下提供目标验证。
Dissecting complex biological systems requires target-specific control of protein function or abundance. Genetic perturbations are limited by off-target effects, multi-component complexity, and irreversibility. Most limiting is the requisite delay from modulation to experimental measurement. To enable the immediate and selective control of single protein abundance, we created a chemical biology system that leverages the potency of cell-permeable heterobifunctional degraders. The dTAG system pairs a novel degrader of FKBP12F36V with expression of FKBP12F36V in-frame with a protein of interest. By transgene expression or CRISPR-mediated locus-specific knock-in, we exemplify a generalizable strategy to study the immediate consequence of protein loss. Using dTAG, we observe an unexpected superior anti-proliferative effect of pan-BET bromodomain degradation over selective BRD4 degradation, characterize immediate effects of KRASG12V loss on proteomic signaling, and demonstrate rapid degradation in vivo. This technology platform will confer kinetic resolution to biological investigation and provide target validation in the context of drug discovery.
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选择性抑制BET溴结构域。
影响因子：
64.8
作者：
通讯作者：
--
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：
16.8
作者：
Deshpande AJ;Bradner J;Armstrong SA
通讯作者：
Armstrong SA
影响因子：
64.5
作者：
Anand P;Brown JD;Lin CY;Qi J;Zhang R;Artero PC;Alaiti MA;Bullard J;Alazem K;Margulies KB;Cappola TP;Lemieux M;Plutzky J;Bradner JE;Haldar SM
通讯作者：
Haldar SM
影响因子：
14.8
作者：
Bondeson DP;Mares A;Smith IE;Ko E;Campos S;Miah AH;Mulholland KE;Routly N;Buckley DL;Gustafson JL;Zinn N;Grandi P;Shimamura S;Bergamini G;Faelth-Savitski M;Bantscheff M;Cox C;Gordon DA;Willard RR;Flanagan JJ;Casillas LN;Votta BJ;den Besten W;Famm K;Kruidenier L;Carter PS;Harling JD;Churcher I;Crews CM
通讯作者：
Crews CM