Exciton-Controlled Hybridization-Sensitive Fluorescent Probes: Multicolor Detection of Nucleic Acids
Exciton-Controlled Hybridization-Sensitive Fluorescent Probes: Multicolor Detection of Nucleic Acids
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DOI：
10.1002/anie.200902000
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发表时间：
2009-01-01
影响因子：
16.6
通讯作者：
Okamoto, Akimitsu
中科院分区：
文献类型：
--
作者：
Ikeda, Shuji;Kubota, Takeshi;Okamoto, Akimitsu
设计一种荧光探针，使其在不识别目标核酸时关闭荧光，对于建立核酸成像方法是非常重要的。核酸检测对荧光探针的要求不仅是序列选择性发射和避免非特异性发射，而且重要的是确保发射是多色的，以同时监测不同的目标。到目前为止，以下内容已被用于包含“开-关”开关系统的高功能荧光探针的分子设计：光物理和光化学(例如，准分子形成、[1]光致电荷转移、[2]光致电子转移、[3]和荧光共振能量转移)。[4]在这些探针的设计中，染料对环境的敏感性、更高有序的探针构象以及通过电子从/向碱基转移而猝灭都对荧光强度和灵敏度有很大影响，并且经常是配对预测。探针的多色性还受到荧光切换机制、探针构象或合成过程的限制。我们重点研究了噻唑橙荧光染料的激子相互作用，并设计了一种新的高效核酸检测方法的双荧光标记核苷酸。[5-7]两个噻唑橙染料分子连接到嘧啶碱基上的探针的荧光很好地受到激子相互作用的控制(图1)。激子相互作用是通过染料之间形成H聚集体而产生的。结果，抑制了杂交前来自探针的发射。
The design of a fluorescent probe in which the fluorescence is switched off when the probe does not recognize the target nucleic acid is very important for the establishment of a nucleic acid imaging method. The requirements for fluorescent probes for nucleic acid detection are not only sequence-selective emission and the avoidance of nonspecific emission, but it is also important to ensure that the emission is polychrome for the simultaneous monitoring of different targets. The following have been used to date for the molecular design of highly functional fluorescent probes containing an “on–off” switching system: photophysics and photochemistry (eg excimer formation,[1] photoinduced charge transfer,[2] photoinduced electron transfer,[3] and fluorescence resonance energy transfer).[4] In the design of these probes, the sensitivity of the dye to the environment, higher-ordered probe conformations, and quenching by electron transfer from/to nucleobases all have a strong influence on fluorescence intensity and sensitivity and often impair predictions. The multicoloring of probes is also limited by a fluorescenceswitching mechanism, the probe conformation, or synthetic processes.We have focused on the excitonic interaction observed for thiazole orange fluorescent dyes and designed a doubly fluorescence labeled nucleotide for a new efficient nucleic acid detection method.[5–7] The fluorescence of the probe in which two thiazole orange dye molecules are attached to a pyrimidine base is well-controlled by an excitonic interaction (Figure1). An excitonic interaction is produced by the formation of an H aggregate between the dyes. As a result, emission from the probe before hybridization is suppressed.