Specific detection of the cleavage activity of mycobacterial enzymes using a quantum dot based DNA nanosensor

Morten Leth Jepsen, Charlotte Harmsen, Adwait Anand Godbole, Valakunja Nagaraja, Birgitta R. Knudsen, Yi Ping Ho*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

We present a quantum dot based DNA nanosensor specifically targeting the cleavage step in the reaction cycle of the essential DNA-modifying enzyme, mycobacterial topoisomerase I. The design takes advantages of the unique photophysical properties of quantum dots to generate visible fluorescence recovery upon specific cleavage by mycobacterial topoisomerase I. This report, for the first time, demonstrates the possibility to quantify the cleavage activity of the mycobacterial enzyme without the pre-processing sample purification or post-processing signal amplification. The cleavage induced signal response has also proven reliable in biological matrices, such as whole cell extracts prepared from Escherichia coli and human Caco-2 cells. It is expected that the assay may contribute to the clinical diagnostics of bacterial diseases, as well as the evaluation of treatment outcomes.
Original languageEnglish
JournalNanoscale
Volume8
Issue number1
Pages (from-to)358-364
ISSN2040-3364
DOIs
Publication statusPublished - 7 Jan 2016
Externally publishedYes

Cite this

Jepsen, Morten Leth ; Harmsen, Charlotte ; Godbole, Adwait Anand ; Nagaraja, Valakunja ; Knudsen, Birgitta R. ; Ho, Yi Ping. / Specific detection of the cleavage activity of mycobacterial enzymes using a quantum dot based DNA nanosensor. In: Nanoscale. 2016 ; Vol. 8, No. 1. pp. 358-364.
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abstract = "We present a quantum dot based DNA nanosensor specifically targeting the cleavage step in the reaction cycle of the essential DNA-modifying enzyme, mycobacterial topoisomerase I. The design takes advantages of the unique photophysical properties of quantum dots to generate visible fluorescence recovery upon specific cleavage by mycobacterial topoisomerase I. This report, for the first time, demonstrates the possibility to quantify the cleavage activity of the mycobacterial enzyme without the pre-processing sample purification or post-processing signal amplification. The cleavage induced signal response has also proven reliable in biological matrices, such as whole cell extracts prepared from Escherichia coli and human Caco-2 cells. It is expected that the assay may contribute to the clinical diagnostics of bacterial diseases, as well as the evaluation of treatment outcomes.",
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Specific detection of the cleavage activity of mycobacterial enzymes using a quantum dot based DNA nanosensor. / Jepsen, Morten Leth; Harmsen, Charlotte; Godbole, Adwait Anand; Nagaraja, Valakunja; Knudsen, Birgitta R.; Ho, Yi Ping.

In: Nanoscale, Vol. 8, No. 1, 07.01.2016, p. 358-364.

Research output: Contribution to journalJournal articleResearchpeer-review

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AU - Jepsen, Morten Leth

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AU - Godbole, Adwait Anand

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AB - We present a quantum dot based DNA nanosensor specifically targeting the cleavage step in the reaction cycle of the essential DNA-modifying enzyme, mycobacterial topoisomerase I. The design takes advantages of the unique photophysical properties of quantum dots to generate visible fluorescence recovery upon specific cleavage by mycobacterial topoisomerase I. This report, for the first time, demonstrates the possibility to quantify the cleavage activity of the mycobacterial enzyme without the pre-processing sample purification or post-processing signal amplification. The cleavage induced signal response has also proven reliable in biological matrices, such as whole cell extracts prepared from Escherichia coli and human Caco-2 cells. It is expected that the assay may contribute to the clinical diagnostics of bacterial diseases, as well as the evaluation of treatment outcomes.

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