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Genetically encoded biosensors for the detection of rapamycin: toward the screening of agonists and antagonists.
Analyst ( IF 3.6 ) Pub Date : 2020-06-18 , DOI: 10.1039/d0an01116a Euiyeon Lee 1 , Hyunjin Jeon , Jeahee Ryu , Chungwon Kang , Soyoun Kim , Seungil Park , Youngeun Kwon
Analyst ( IF 3.6 ) Pub Date : 2020-06-18 , DOI: 10.1039/d0an01116a Euiyeon Lee 1 , Hyunjin Jeon , Jeahee Ryu , Chungwon Kang , Soyoun Kim , Seungil Park , Youngeun Kwon
Affiliation
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Biosensors are valuable tools for the rapid screening of biological targets with high sensitivity and specificity. It is important to screen biological events in their native context for pharmacological and toxicological applications. However, in vitro biosensors often require purified probes and targets for screening, thus providing limited information on the biological activities of targets in their native environment. To address this issue, we developed a cell-based sensing system that could detect a biologically active small molecule, rapamycin (Rapa). We designed a reporter system based on fluorescence translocation by signal peptide reconstitution. Herein, signal peptides are activated by conditional protein splicing without the need for refolding into a functional tertiary structure, thus eliminating false positives and negatives due to mere binding or misfolding. The developed biosensor demonstrated excellent sensitivity with a limit of detection of 0.1 nM, and it was able to screen the agonist and antagonist of Rapa. The developed cell-based sensing system could contribute to improving the screening system aimed to identify the natural mimetics of Rapa and potential drug candidates.
中文翻译：
用于雷帕霉素检测的遗传编码生物传感器：用于激动剂和拮抗剂的筛选。
生物传感器是用于以高灵敏度和特异性快速筛选生物靶标的有价值的工具。对于药物和毒理学应用，在其自然环境中筛选生物事件非常重要。但是，体外生物传感器通常需要纯化的探针和靶标进行筛选，因此提供了有关靶标在其天然环境中的生物学活性的有限信息。为了解决这个问题，我们开发了一种基于细胞的传感系统，可以检测具有生物活性的小分子雷帕霉素（Rapa）。我们设计了一种基于信号肽重构的荧光易位的报告系统。在此，信号肽通过条件蛋白剪接而被激活，而不需要重新折叠成功能性三级结构，从而消除了仅仅由于结合或错误折叠而引起的假阳性和假阴性。研发的生物传感器显示出极好的灵敏度，检测限为0.1 nM，并且能够筛选Rapa的激动剂和拮抗剂。
更新日期：2020-08-10
中文翻译：
用于雷帕霉素检测的遗传编码生物传感器：用于激动剂和拮抗剂的筛选。
生物传感器是用于以高灵敏度和特异性快速筛选生物靶标的有价值的工具。对于药物和毒理学应用，在其自然环境中筛选生物事件非常重要。但是，体外生物传感器通常需要纯化的探针和靶标进行筛选，因此提供了有关靶标在其天然环境中的生物学活性的有限信息。为了解决这个问题，我们开发了一种基于细胞的传感系统，可以检测具有生物活性的小分子雷帕霉素（Rapa）。我们设计了一种基于信号肽重构的荧光易位的报告系统。在此，信号肽通过条件蛋白剪接而被激活，而不需要重新折叠成功能性三级结构，从而消除了仅仅由于结合或错误折叠而引起的假阳性和假阴性。研发的生物传感器显示出极好的灵敏度，检测限为0.1 nM，并且能够筛选Rapa的激动剂和拮抗剂。
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