Document Type

Journal Article

Department/Unit

Department of Chemistry

Title

Detection of nicking endonuclease activity using a G-quadruplex-selective luminescent switch-on probe

Language

English

Abstract

© the Partner Organisations 2014. A series of luminescent Ir(iii) complexes were synthesised and evaluated for their ability to act as G-quadruplex-selective probes. A novel Ir(iii) complex was found to be highly selective for G-quadruplex DNA, and was employed in a label-free G-quadruplex-based detection assay for nicking endonuclease activity in aqueous solution. A proof-of-concept of this probe has been demonstrated by using Nb·BbvCI as a model enzyme. In this assay, a DNA substrate comprised of oligonucleotides ON1 (5′-GTG3TAG3CG3T2G2CTGAG2TGA-3′) and ON2 (5′-TCAC2TCAGC2A2C2-3′) initially exists in a duplex conformation, resulting in a low luminescence signal due to the weak interaction between the Ir(iii) complex and duplex DNA. Upon cleavage by Nb·BbvCI, the guanine-rich sequence is released and folds into a G-quadruplex, which greatly enhances the luminescence of the Ir(iii) probe. This method was highly sensitive for Nb·BbvCI over other DNA-modifying enzymes. This journal is

Publication Date

2014

Source Publication Title

Chemical Science

Volume

5

Issue

12

Start Page

4561

End Page

4568

Publisher

Royal Society of Chemistry

DOI

10.1039/C4SC02032D

Link to Publisher's Edition

http://dx.doi.org/10.1039/C4SC02032D

ISSN (print)

20416520

ISSN (electronic)

20416539

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