A Förster Resonance Energy Transfer-Based Ratiometric Sensor with the Allosteric Transcription Factor TetR.,Small

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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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