Label-free Molecular Beacons for Biomolecular Detection

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• 作者：Xiaohong Tan ; Yi Wang ; Bruce A. Armitage ; Marcel P. Bruchez
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：November 4, 2014
• 年：2014
• 卷：86
• 期：21
• 页码：10864-10869
• 全文大小：321K
• ISSN：1520-6882

文摘

Biomolecular detection and imaging methods provide quantitative measurements essential for biological research. In this context, molecular beacon based sensors have emerged as powerful, no-wash imaging agents, providing target-specific fluorescent activation for nucleic acids, proteins, and small molecules. Conventional molecular beacons require double-labeled DNA sequences, which are costly and time-consuming to prepare. To address this issue, we developed DNA based label-free molecular beacons consisting of two regions: a signal-generating region based on human telomeric G-quadruplex sequence that activates Thioflavin T fluorescence and a target recognition sequence designed to interact in a molecular beacon format. We demonstrated the utility of these probes for the selective detection of DNA, RNA, and protein. Multiple probes were applied against a single target to achieve improved brightness in fluorescence detection of nucleic acid targets. This label-free strategy provides a straightforward, cost-effective alternative to fluorescently labeled oligonucleotides in biomolecular detection and imaging.