Conditional Protein Rescue by Binding-Induced Protective Shielding,ACS Synthetic Biology

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Conditional Protein Rescue by Binding-Induced Protective Shielding
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-10-07 , DOI: 10.1021/acssynbio.0c00367
Andrew S Gaynor ₁ , Wilfred Chen ₁

Affiliation

Synthetic protein-level circuits offer an extra layer of cellular control on top of conventional gene-level circuits. Here, we describe a technology that allows conditional protein rescue (CPR) from proteasomal degradation using different protein inputs as masking agents. A target protein is fused to a degron tag and an affinity sensor domain. The use of nanobodies as the sensor domain offers a generalizable strategy to execute a wide range of protein-level circuits with ease. The utility of this new strategy was successfully demonstrated to distinguish cancer cells out of a healthy population using the HPV-specific E7 protein as a cellular marker. Because CPR can be programmed to execute more complex Boolean logic designs using cell-specific proteomes, this platform offers a highly modular and scalable framework for a wide range of applications based on synthetic protein circuits.

中文翻译：

通过结合诱导的保护性屏蔽来有条件地拯救蛋白质

合成蛋白质水平电路在传统基因水平电路之上提供了一层额外的细胞控制。在这里，我们描述了一种技术，该技术允许使用不同的蛋白质输入作为掩蔽剂从蛋白酶体降解中进行条件蛋白质拯救 (CPR)。目标蛋白与 degron 标签和亲和传感器域融合。使用纳米抗体作为传感器域提供了一种通用策略，可以轻松执行各种蛋白质级电路。使用 HPV 特异性 E7 蛋白作为细胞标记物，成功地证明了这种新策略的效用可以将癌细胞与健康人群区分开来。因为 CPR 可以被编程为使用细胞特异性蛋白质组执行更复杂的布尔逻辑设计，

更新日期：2020-10-17

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