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A Förster Resonance Energy Transfer-Based Ratiometric Sensor with the Allosteric Transcription Factor TetR.
Small ( IF 13.0 ) Pub Date : 2020-04-06 , DOI: 10.1002/smll.201907522
Thuy T Nguyen 1 , Margaret Chern 2 , R C Baer 3 , James Galagan 1, 3, 4 , Allison M Dennis 1, 2
Affiliation  

A recent description of an antibody-free assay is significantly extended for small molecule analytes using allosteric transcription factors (aTFs) and Förster resonance energy transfer (FRET). The FRET signal indicates the differential binding of an aTF-DNA pair with a dose-dependent response to its effector molecule, i.e., the analyte. The new sensors described here, based on the well-characterized aTF TetR, demonstrate several new features of the assay approach: 1) the generalizability of the sensors to additional aTF-DNA-analyte systems, 2) sensitivity modulation through the choice of donor fluorophore (quantum dots or fluorescent proteins, FPs), and 3) sensor tuning using aTF variants with differing aTF-DNA binding affinities. While all of these modular sensors self-assemble, the design reported here based on a recombinant aTF-FP chimera with commercially available dye-labeled DNA uses readily accessible sensor components to facilitate easy adoption of the sensing approach by the broader community.

中文翻译:


具有变构转录因子 TetR 的基于福斯特共振能量转移的比例传感器。



最近对无抗体测定的描述显着扩展到使用变构转录因子 (aTF) 和福斯特共振能量转移 (FRET) 的小分子分析物。 FRET 信号表明 aTF-DNA 对的差异结合与其效应分子(即分析物)具有剂量依赖性响应。这里描述的新传感器基于充分表征的 aTF TetR,展示了该检测方法的几个新特征:1) 传感器对其他 aTF-DNA 分析物系统的通用性,2) 通过选择供体荧光团进行灵敏度调节(量子点或荧光蛋白,FP),3) 使用具有不同 aTF-DNA 结合亲和力的 aTF 变体进行传感器调谐。虽然所有这些模块化传感器都是自组装的,但本文报告的设计基于重组 aTF-FP 嵌合体和市售染料标记 DNA,使用易于访问的传感器组件,以促进更广泛的社区轻松采用传感方法。
更新日期:2020-04-06
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