Sensors 2013, 13(4), 4017-4028; doi:10.3390/s130404017
Article

DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I

Received: 1 February 2013; in revised form: 16 February 2013 / Accepted: 19 March 2013 / Published: 25 March 2013
(This article belongs to the Special Issue Enzymatic Biosensors)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Sensors capable of quantitative real-time measurements may present the easiest and most accurate way to study enzyme activities. Here we present a novel DNA-based sensor for specific and quantitative real-time measurement of the enzymatic activity of the essential human enzyme, topoisomerase I. The basic design of the sensor relies on two DNA strands that hybridize to form a hairpin structure with a fluorophore-quencher pair. The quencher moiety is released from the sensor upon reaction with human topoisomerase I thus enabling real-time optical measurement of enzymatic activity. The sensor is specific for topoisomerase I even in raw cell extracts and presents a simple mean of following enzyme kinetics using standard laboratory equipment such as a qPCR machine or fluorimeter. Human topoisomerase I is a well-known target for the clinically used anti-cancer drugs of the camptothecin family. The cytotoxic effect of camptothecins correlates directly with the intracellular topoisomerase I activity. We therefore envision that the presented sensor may find use for the prediction of cellular drug response. Moreover, inhibition of topoisomerase I by camptothecin is readily detectable using the presented DNA sensor, suggesting a potential application of the sensor for first line screening for potential topoisomerase I targeting anti-cancer drugs.
Keywords: real-time activity measurement; DNA sensor; fluorophore-quencher pair; topoisomerase I; cancer treatment; camptothecin
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MDPI and ACS Style

Marcussen, L.B.; Jepsen, M.L.; Kristoffersen, E.L.; Franch, O.; Proszek, J.; Ho, Y.-P.; Stougaard, M.; Knudsen, B.R. DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I. Sensors 2013, 13, 4017-4028.

AMA Style

Marcussen LB, Jepsen ML, Kristoffersen EL, Franch O, Proszek J, Ho Y-P, Stougaard M, Knudsen BR. DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I. Sensors. 2013; 13(4):4017-4028.

Chicago/Turabian Style

Marcussen, Lærke B.; Jepsen, Morten L.; Kristoffersen, Emil L.; Franch, Oskar; Proszek, Joanna; Ho, Yi-Ping; Stougaard, Magnus; Knudsen, Birgitta R. 2013. "DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I." Sensors 13, no. 4: 4017-4028.

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