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基于分子间 G-四链体结构的荧光 DNA 检测体系。

Intermolecular G-quadruplex structure-based fluorescent DNA detection system.

机构信息

State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.

出版信息

Biosens Bioelectron. 2013 Mar 15;41:262-7. doi: 10.1016/j.bios.2012.08.028. Epub 2012 Aug 31.

Abstract

Adopting multi-donors to pair with one acceptor could improve the performance of fluorogenic detection probes. However, common dyes (e.g., fluorescein) in close proximity to each other would self-quench the fluorescence, and the fluorescence is difficult to restore. In this contribution, we constructed a novel "multi-donors-to-one acceptor" fluorescent DNA detection system by means of the intermolecular G-quadruplex (IGQ) structure-based fluorescence signal enhancement combined with the hairpin oligonucleotide. The novel IGQ-hairpin system was characterized using the p53 gene as the model target DNA. The proposed system showed an improved assay performance due to the introduction of IGQ-structure into fluorescent signaling probes, which could inhibit the background fluorescence and increase fluorescence restoration amplitude of fluoresceins upon target DNA hybridization. The proof-of-concept scheme is expected to provide new insight into the potential of G-quadruplex structure and promote the application of fluorescent oligonucleotide probes in fundamental research, diagnosis, and treatment of genetic diseases.

摘要

采用多供体与一个受体配对可以提高荧光检测探针的性能。然而,彼此靠近的常见染料(例如荧光素)会自猝灭荧光,并且荧光难以恢复。在本研究中,我们通过基于分子间 G-四链体(IGQ)结构的荧光信号增强与发夹寡核苷酸相结合,构建了一种新型的“多供体到一个受体”荧光 DNA 检测系统。新型 IGQ-发夹系统以 p53 基因为模型靶 DNA 进行了表征。由于将 IGQ 结构引入到荧光信号探针中,该系统的检测性能得到了改善,这可以抑制背景荧光并增加靶 DNA 杂交时荧光素的荧光恢复幅度。这一概念验证方案有望为 G-四链体结构的潜力提供新的见解,并促进荧光寡核苷酸探针在基础研究、诊断和遗传疾病治疗中的应用。

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