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Simple, PCR-free telomerase activity detection using G-quadruplex–hemin DNAzyme


Hui Li, Deming Kong et al.


RSC Adv., 2015, 5, 6475


A simple, cost-effective and polymerase chain reaction (PCR)-free telomerase activity detection method was developed on the basis of telomerase-triggered formation of G-quadruplex-hemin DNAzyme. In this method, a short, unlabelled telomerase primer was used. Because this primer contains only three GGG repeats, it cannot fold into the stable G-quadruplex structure. In the presence of active telomerase and dGTP, a GGG repeat is added to the 3'-end of the primer. The extended primer can fold into the G-quadruplex, which is able to bind hemin to form catalytically active G-quadruplex-hemin DNAzyme, catalyzing the oxidation of 2,2'-azinobis (3-ethylbenzothiozoline)-6-sulfonic acid (ABTS) by H2O2 to green ABTSc(center dot+). Because the primer extension product is very short, telomerase should show a high turnover rate, thus providing the method with improved sensitivity. Using this method, the telomerase activity originating from 200 HeLa cells can be detected.