College of Chemistry, Nanchang University, Nanchang, 330031, China.
Department of Materials and Chemical Engineering, Pingxiang University, 211 Pingan North Road, Pingxiang, 337055, Jiangxi, China.
Mikrochim Acta. 2019 Apr 29;186(5):309. doi: 10.1007/s00604-019-3391-z.
The authors describe cobalt phosphide (CoP) nanowires for use in sensitive fluorometric determination of the activity of the enzyme telomerase. A hybridization chain reaction (HCR) is applied to amplify the signal and carboxyfluorescein (FAM)-labelled hairpin probes (H1 and H2) are applied to match the telomeric DNA sequence. The CoP nanowires act as both the photoinduced electron transfer (PET) acceptor to induce fluorescence quenching, and also as an efficient probe carrier to facilitate telomerase imaging in living cells. The telomerase-triggered primer extension initiates an alternating hybridization reaction between H1 and H2. These result in the dissociation of FAM-labelled probes from CoP nanowires and thus an enhancement of the green fluorescence. The method is fairly simple and was applied to the detection of three types of cancer cells. The detection limit is as low as 7 cells (in case of HeLa cells). Conceivably, the method has a large potential in terms of inhibitor drug screening. Graphical abstract Schematic presentation of telomerase detection based on cobalt phosphide (CoP) nanowires and hybridization chain reaction (HCR). The telomerase-triggered primer extension can initiate the alternating hybridization reaction between carboxyfluorescein (FAM)-labelled hairpin probes (H1 and H2), and the generated long DNA duplex cannot be adsorbed on the CoP nanowires. This prevents the photoinduced electron transfer (PET) from FAM to CoP nanowires.
作者描述了用于灵敏荧光测定端粒酶活性的磷化钴(CoP)纳米线。应用杂交链式反应(HCR)进行信号放大,并用羧基荧光素(FAM)标记的发夹探针(H1 和 H2)匹配端粒 DNA 序列。CoP 纳米线既可以作为光诱导电子转移(PET)的受体来诱导荧光猝灭,也可以作为有效的探针载体,促进端粒酶在活细胞中的成像。端粒酶触发的引物延伸引发 H1 和 H2 之间的交替杂交反应。这导致 FAM 标记的探针从 CoP 纳米线解离,从而增强绿色荧光。该方法相当简单,并应用于三种类型的癌细胞的检测。检测限低至 7 个细胞(以 HeLa 细胞为例)。可以想象,该方法在抑制剂药物筛选方面具有很大的潜力。示意图基于磷化钴(CoP)纳米线和杂交链式反应(HCR)的端粒酶检测示意图。端粒酶触发的引物延伸可以引发羧基荧光素(FAM)标记的发夹探针(H1 和 H2)之间的交替杂交反应,生成的长 DNA 双链不能被吸附到 CoP 纳米线上。这阻止了 FAM 到 CoP 纳米线的光诱导电子转移(PET)。