A Robust Probe for Lighting Up Intracellular Telomerase via Primer Extension To Open a Nicked Molecular Beacon,Journal of the American Chemical Society

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A Robust Probe for Lighting Up Intracellular Telomerase via Primer Extension To Open a Nicked Molecular Beacon
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2014-05-29 , DOI: 10.1021/ja5042995
Ruocan Qian ₁ , Lin Ding ₁ , Liwen Yan ₁ , Manfei Lin ₁ , Huangxian Ju ₁

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A nicked molecular beacon (MB)-functionalized probe has been designed for in situ imaging and detection of intracellular telomerase activity. The nick separates the MB into two segments: a shorter telomerase primer (TSP) sequence as a part of the 5'-end stem and a longer sequence to form a loop with one thiol-labeled 3'-end stem. The MB can be opened by substitutional hybridization of the telomerase-triggered stem elongation product, which leads to separation of the Cy5 at the 5'-end nick from the gold nanoparticle (AuNP) as the nanocarrier and thus inhibits the energy transfer from Cy5 to AuNP. Upon endocytosis of the probe, the TSP can be extended by intracellular telomerase at its 3' end to produce the telomeric repeated sequence, which leads to the inner chain substitution and thus turns on the fluorescence of Cy5. The probe provides a one-step incubation technique for quantification and monitoring of the telomerase activity in living cells. The practicality of the proposed approach for distinguishing tumor cells from normal cells and monitoring the decrease of telomerase activity during treatment with antitumor drugs demonstrates its potential in clinical diagnostic and therapeutic monitoring.

中文翻译：

通过引物延伸点亮细胞内端粒酶以打开带缺口的分子信标的稳健探针

设计了一种带切口的分子信标 (MB) 功能化探针，用于原位成像和检测细胞内端粒酶活性。缺口将 MB 分成两个部分：较短的端粒酶引物 (TSP) 序列作为 5' 端茎的一部分，较长的序列与一个硫醇标记的 3' 端茎形成环。MB 可以通过端粒酶触发的茎伸长产物的置换杂交打开，这导致 Cy5 在 5' 端切口处与作为纳米载体的金纳米颗粒 (AuNP) 分离，从而抑制能量从 Cy5 转移到金纳米粒子。探针内吞后，TSP 可被胞内端粒酶在其 3' 端延伸，产生端粒重复序列，导致内链取代，从而开启 Cy5 的荧光。该探针提供了一种用于定量和监测活细胞中端粒酶活性的一步孵育技术。所提出的用于区分肿瘤细胞与正常细胞并监测抗肿瘤药物治疗期间端粒酶活性降低的方法的实用性证明了其在临床诊断和治疗监测中的潜力。

更新日期：2014-05-29

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