A Targeted, Self-Delivered, and Photocontrolled Molecular Beacon for mRNA Detection in Living Cells

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• 作者：Liping Qiu ; Cuichen Wu ; Mingxu You ; Da Han ; Tao Chen ; Guizhi Zhu ; Jianhui Jiang ; Ruqin Yu ; Weihong Tan
• 刊名：Journal of the American Chemical Society
• 出版年：2013
• 出版时间：September 4, 2013
• 年：2013
• 卷：135
• 期：35
• 页码：12952-12955
• 全文大小：253K
• 年卷期：v.135,no.35(September 4, 2013)
• ISSN：1520-5126

文摘

The spatiotemporal dynamics of specific mRNA molecules are difficult to image and detect inside living cells, and this has been a significant challenge for the chemical and biomedical communities. To solve this problem, we have developed a targeted, self-delivered, and photocontrolled aptamer-based molecular beacon (MB) for intracellular mRNA analysis. An internalizing aptamer connected via a double-stranded DNA structure was used as a carrier probe (CP) for cell-specific delivery of the MB designed to signal target mRNA. A light activation strategy was employed by inserting two photolabile groups in the CP sequence, enabling control over the MB鈥檚 intracellular function. After the probe was guided to the target cell via specific binding of aptamer AS1411 to nucleolin on the cell membrane, light illumination released the MB for mRNA monitoring. Consequently, the MB is able to perform live-cell mRNA imaging with precise spatiotemporal control, while the CP acts as both a tracer for intracellular distribution of the MB before photoinitiation and an internal reference for mRNA ratiometric detection.