Smart Vesicle Kit for In Situ Monitoring of Intracellular Telomerase Activity Using a Telomerase-Responsive Probe
Smart Vesicle Kit for In Situ Monitoring of Intracellular Telomerase Activity Using a Telomerase-Responsive Probe
复制标题
使用端粒酶响应探针原位监测细胞内端粒酶活性的智能囊泡试剂盒
DOI：
10.1021/ac502538w
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发表时间：
2014-09-02
影响因子：
7.4
通讯作者：
Ju, Huangxian
中科院分区：
文献类型：
--
作者：
Qian, Ruocan;Ding, Lin;Ju, Huangxian
设计了一种用于细胞质端粒酶活性原位成像和检测的智能囊泡试剂盒。囊泡试剂盒包含端粒酶引物（TSP）和cy5标记的分子信标（MB）功能化金纳米粒子探针，它们被包封在脂质体中用于细胞内递送。将囊泡试剂盒转染细胞质后，释放出的TSP在端粒酶存在下可被延长，在3′端产生端粒重复序列，与组装在探针表面的MB环恰好互补。因此，在杂交时打开MB，将荧光状态从“关”切换到“开”。荧光信号依赖于端粒酶活性，从而为细胞质端粒酶活性的原位成像和定量检测提供了一种新的策略。估计每个HeLa、BEL肿瘤和QSG正常细胞的细胞质端粒酶活性分别为3.2 × 10（-11）、2.4 × 10（-11）和8.6 × 10(-13) IU，表明该方法具有区分肿瘤和正常细胞的能力。该方法可用于端粒酶药物对细胞质端粒酶活性的动态监测，在端粒酶靶向抗癌药物的发现和筛选中具有潜在的应用价值。
A smart vesicle kit was designed for in situ imaging and detection of cytoplasmic telomerase activity. The vesicle kit contained a telomerase primer (TSP) and a Cy5-tagged molecular beacon (MB) functionalized gold nanoparticle probe, which were encapsulated in liposome for intracellular delivery. After the vesicle kit was transfected into cytoplasm, the released TSP could be extended in the presence of telomerase to produce a telomeric repeated sequence at the 3' end, which was just complementary with the loop of MB assembled on probe surface. Thus, the MB was opened upon hybridization to switch the fluorescent state from "off" to "on". The fluorescence signal depended on telomerase activity, leading to a novel strategy for in situ imaging and quantitative detection of the cytoplasmic telomerase activity. The cytoplasmic telomerase activity was estimated to be 3.2 x 10(-11), 2.4 x 10(-11), and 8.6 x 10(-13) IU in each HeLa, BEL tumor and QSG normal cell, respectively, demonstrating the capability of this approach to distinguish tumor from normal cells. The proposed method could be employed for dynamic monitoring of the cytoplasmic telomerase activity in response to a telomerase-based drug, suggesting the potential application in discovery and screening of telomerase-targeted anticancer drugs.