Wang Wenjing, Huang Shan, Li Jingjing, Rui Kai, Bi Sai, Zhang Jian-Rong, Zhu Jun-Jie
State Key Laboratory of Analytical Chemistry for Life Science and Collaborative Innovation Center of Chemistry for Life Sciences , School of Chemistry and Chemical Engineering , Nanjing University , Nanjing 210093 , China . Email:
School of Pharmacy , Ningxia Medical University , Yinchuan 750004 , China.
Chem Sci. 2017 Jan 1;8(1):174-180. doi: 10.1039/c6sc01953f. Epub 2016 Aug 1.
Telomerase plays a vital role in cancer and aging, and telomerase activity detection has drawn great attention recently. However, a feasible imaging system for intracellular telomerase is still a challenge. Here, we develop a novel approach to image intracellular telomerase activity using DNA-based computation. A cascade nucleic acid logic gate that responded to intracellular telomerase was constructed. A telomerase substrate (TS) probe, extended by intracellular telomerase, worked as an input to initiate computation cascades. In this way, intracellular telomerase could be clearly indicated by fluorophore labeled nucleic acids as the output. Through one-step incubation, evaluation of the intracellular telomerase activity for a HeLa cell line and the ability to differentiate cancer cells from normal cells could be realized. Furthermore, the response of intracellular telomerase activity to a telomerase-inhibiting model drug was observed using the proposed method. Thus, this intracellular telomerase computation device will allow improvements in studying the relationship between telomerase and cancer, and may help to develop telomerase inhibitors. This finding also expands the applications of DNA computational techniques in cells.
端粒酶在癌症和衰老过程中起着至关重要的作用,近年来端粒酶活性检测备受关注。然而,构建一种可行的细胞内端粒酶成像系统仍是一项挑战。在此,我们开发了一种基于DNA计算的新方法来对细胞内端粒酶活性进行成像。构建了一种对细胞内端粒酶有响应的级联核酸逻辑门。由细胞内端粒酶延伸的端粒酶底物(TS)探针作为输入启动计算级联反应。通过这种方式,细胞内端粒酶可以通过荧光团标记的核酸作为输出清晰地显示出来。经过一步孵育,可实现对HeLa细胞系细胞内端粒酶活性的评估以及区分癌细胞和正常细胞的能力。此外,使用所提出的方法观察了细胞内端粒酶活性对端粒酶抑制模型药物的反应。因此,这种细胞内端粒酶计算装置将有助于改进对端粒酶与癌症之间关系的研究,并可能有助于开发端粒酶抑制剂。这一发现也扩展了DNA计算技术在细胞中的应用。