Dynamic tracing of intracellular telomerase activity plays a crucial role in cancer cell recognition and correspondingly in earlier cancer diagnosis and personalized precision therapy. However, due to the complexity of the required reaction system and insufficient loading of reaction components into cells, achieving a high-fidelity determination of telomerase activity is still a challenge. Herein, an Aptamer-Liposome mediated Telomerase activated poly-Molecular beacon Arborescent Nanoassembly(ALTMAN) approach was described for direct high-fidelity visualization of telomerase activity. Briefly, intracellular telomerase activates molecular beacons, causing their hairpin structures to unfold and produce fluorescent signals. Furthermore, multiple molecular beacons can self-assemble, forming arborescent nanostructures and leading to exponential amplification of fluorescent signals. Integrating the enzyme-free isothermal signal amplification successfully increased the sensitivity and reduced interference by leveraging the skillful design of the molecular beacon and the extension of the telomerase-activated TTAGGG repeat sequence. The proposed approach enabled ultrasensitive visualization of activated telomerase exclusively with a prominent detection limit of 2 cells·μL^–1 and realized real-time imaging of telomerase activity in living cancer cells including blood samples from breast cancer patients and urine samples from bladder cancer patients. This approach opens an avenue for establishing a telomerase activity determination and in situ monitoring technique that can facilitate both telomerase fundamental biological studies and cancer diagnostics.

中文翻译：

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高保真灵敏追踪循环肿瘤细胞端粒酶活性

细胞内端粒酶活性的动态追踪在癌细胞识别以及相应的早期癌症诊断和个性化精准治疗中起着至关重要的作用。然而，由于所需反应系统的复杂性以及反应组分加载到细胞中的不足，实现端粒酶活性的高保真度测定仍然是一个挑战。在此，描述了适体-脂质体介导的端粒酶激活的多分子信标树状纳米组装（ALTMAN）方法，用于端粒酶活性的直接高保真可视化。简而言之，细胞内端粒酶激活分子信标，导致其发夹结构展开并产生荧光信号。此外，多个分子信标可以自组装，形成树状纳米结构并导致荧光信号指数放大。通过利用分子信标的巧妙设计和端粒酶激活的 TTAGGG 重复序列的延伸，集成无酶等温信号放大成功地提高了灵敏度并减少了干扰。该方法能够以2个细胞·μL ^–1的显着检测限实现激活端粒酶的超灵敏可视化，并实现活体癌细胞中端粒酶活性的实时成像，包括乳腺癌患者的血液样本和膀胱癌患者的尿液样本。这种方法为建立端粒酶活性测定和原位监测技术开辟了一条途径，可以促进端粒酶基础生物学研究和癌症诊断。