DNA-based sensor for real-time measurement of the enzymatic activity of human topoisomerase I.
Sensors (Basel)
; 13(4): 4017-28, 2013 Mar 25.
Article
em En
| MEDLINE
| ID: mdl-23529147
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sistemas Computacionais
/
DNA
/
Técnicas Biossensoriais
/
DNA Topoisomerases Tipo I
Limite:
Humans
Idioma:
En
Revista:
Sensors (Basel)
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Dinamarca