Issue 19, 2018

A simple “turn-on” fluorescent biosensor for sensitive detection of exonuclease III activity through photoinduced electron transfer and self-hybridization of a DNA probe

Abstract

Exonuclease III (Exo III) plays crucial roles in maintaining the genome stability. Herein, a turn-on fluorescence strategy for detection of Exo III activity is developed. In this strategy, a poly(CG) single-stranded DNA (ssDNA) probe is labeled with fluorescein amidite (FAM) at the 3′ end, and the ssDNA can self-hybridize to form double-stranded DNA (dsDNA). This dsDNA acts as the substrate for Exo III. Due to photoinduced electron transfer (PET) between FAM and guanine (G), the fluorescence of the FAM-labeled dsDNA probe is completely quenched. Upon the addition of Exo III, the dsDNA will be digested, and FAM emits very strong fluorescence. Thus, Exo III activity can be facilely measured with a simple fluorescence reader. This method has a wide detection range from 5 × 10−4 U mL−1 to 5 U mL−1 with a detection limit of 3 × 10−4 U mL−1. The results demonstrated herein may provide a new pattern for Exo III activity detection with high accuracy and good specificity as well as satisfactory applicability in real biosamples, which holds great potential for drug screening and basic research related to exonucleases.

Graphical abstract: A simple “turn-on” fluorescent biosensor for sensitive detection of exonuclease III activity through photoinduced electron transfer and self-hybridization of a DNA probe

Supplementary files

Article information

Article type
Paper
Submitted
22 Mar 2018
Accepted
09 Apr 2018
First published
10 Apr 2018

Anal. Methods, 2018,10, 2257-2262

A simple “turn-on” fluorescent biosensor for sensitive detection of exonuclease III activity through photoinduced electron transfer and self-hybridization of a DNA probe

Q. Liu, J. Lian, M. Liu, Y. Jin and B. Li, Anal. Methods, 2018, 10, 2257 DOI: 10.1039/C8AY00631H

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements