Immobilization of Cyanines in DNA Produces Systematic Increases in Fluorescence Intensity,The Journal of Physical Chemistry Letters

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Immobilization of Cyanines in DNA Produces Systematic Increases in Fluorescence Intensity
The Journal of Physical Chemistry Letters ( IF 4.8 ) Pub Date : 2021-09-10 , DOI: 10.1021/acs.jpclett.1c02022
Natalie A Pace ₁ , Scott P Hennelly ₂ , Peter M Goodwin ₁

Affiliation

Cyanines are useful fluorophores for a myriad of biological labeling applications, but their interactions with biomolecules are unpredictable. Cyanine fluorescence intensity can be highly variable due to complex photoisomerization kinetics, which are exceedingly sensitive to the surrounding environment. This introduces large errors in Förster resonance energy transfer (FRET)-based experiments where fluorescence intensity is the output parameter. However, this environmental sensitivity is a strength from a biological sensing point of view if specific relationships between biomolecular structure and cyanine photophysics can be identified. We describe a set of DNA structures that modulate cyanine fluorescence intensity through the insertion of adenine or thymine bases. These structures simultaneously provide photophysical predictability and tunability. We characterize these structures using steady-state fluorescence measurements, fluorescence correlation spectroscopy (FCS), and time-resolved photoluminescence (TRPL). We find that the photoisomerization rate decreases over an order of magnitude across the adenine series, which is consistent with increasing immobilization of the cyanine moiety by the surrounding DNA structure.

中文翻译：

将花青固定在 DNA 中会导致荧光强度的系统性增加

花青是用于无数生物标记应用的有用荧光团，但它们与生物分子的相互作用是不可预测的。由于对周围环境极其敏感的复杂光异构化动力学，花青荧光强度可能会发生很大变化。这在基于 Förster 共振能量转移 (FRET) 的实验中引入了大误差，其中荧光强度是输出参数。然而，如果可以确定生物分子结构和花青光物理之间的特定关系，那么从生物传感的角度来看，这种环境敏感性是一种优势。我们描述了一组通过插入腺嘌呤或胸腺嘧啶碱基来调节花青荧光强度的 DNA 结构。这些结构同时提供光物理可预测性和可调性。我们使用稳态荧光测量、荧光相关光谱 (FCS) 和时间分辨光致发光 (TRPL) 来表征这些结构。我们发现光异构化速率在整个腺嘌呤系列中降低了一个数量级，这与周围 DNA 结构对花青部分的固定化增加一致。

更新日期：2021-09-23

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