Pu Fang, Ren Jinsong, Qu Xiaogang
Laboratory of Chemical Biology and State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, China.
Methods Mol Biol. 2019;2035:347-356. doi: 10.1007/978-1-4939-9666-7_21.
G-quadruplexes formed by 3'-overhang of guanine-rich human telomeric DNA at the end of chromosome have important implication in inhibiting the telomerase activity. Telomerase catalyzes the elongation of telomeres by adding telomeric repeats sequence TTAGGG onto the end of the chromosome. Since telomerase is over-expressed in 80-90% of all known human tumors, the enzyme can be recognized as a biomarker for cancer diagnosis and a therapeutic target. Thus, the sensitive detection of telomerase activity is essential to cancer diagnosis and therapy, and screening of anticancer drugs. Gold nanoparticles (AuNPs) have been widely applied as a colorimetric probe for assay owing to their unique size- and distance-dependent optical properties. Human telomerase activity can be visualized by using primer-modified Au nanoparticles. The extremely high extinction coefficients of AuNPs offered high sensitivity. Here, we describe a protocol for the preparation of primer-modified Au nanoparticles for colorimetric assay of human telomerase activity and initial screening of telomerase inhibitors.
由富含鸟嘌呤的人类端粒DNA在染色体末端形成的3'-突出端G-四链体在抑制端粒酶活性方面具有重要意义。端粒酶通过将端粒重复序列TTAGGG添加到染色体末端来催化端粒的延长。由于端粒酶在所有已知人类肿瘤的80%-90%中过度表达，该酶可被视为癌症诊断的生物标志物和治疗靶点。因此，端粒酶活性的灵敏检测对于癌症诊断、治疗以及抗癌药物筛选至关重要。金纳米颗粒（AuNPs）因其独特的尺寸和距离依赖性光学性质而被广泛用作比色测定探针。通过使用引物修饰的金纳米颗粒可以直观地检测人类端粒酶活性。AuNPs极高的消光系数提供了高灵敏度。在此，我们描述了一种制备引物修饰的金纳米颗粒的方法，用于人类端粒酶活性的比色测定和端粒酶抑制剂的初步筛选。
