Issue 22, 2022

Ultrasensitive detection of DNA methyltransferase activity: a novel dual-amplification fluorescence technique

Abstract

DNA methyltransferase (MTase) is an important regulatory enzyme in various biological processes. However, current methods for investigating MTase activity are still limited in terms of sensitivity and/or generality. Herein, we proposed a dual amplification fluorescence strategy for the ultrasensitive detection of DNA adenine methylation methyltransferase (Dam MTase) activity based on strand displacement amplification (SDA) coupled with rolling circle amplification (RCA). In this study, the hairpin probe could not be cleaved by Nt.AlwI nicking endonuclease (Nt.AlwI) in the presence of Dam MTase, and the subsequent SDA–RCA reaction was blocked, resulting in a weak fluorescence signal. Moreover, the blocking effect was more pronounced at a higher concentration of Dam MTase. This assay provides a very low detection limit (down to 0.0067 U ml−1), as well as good selectivity against other types of MTases (e.g., CpG methyltransferase (M.SssI MTase)). In addition, the analytical mode improves the generality and can be extended to the detection of other types of DNA MTases.

Graphical abstract: Ultrasensitive detection of DNA methyltransferase activity: a novel dual-amplification fluorescence technique

Article information

Article type
Paper
Submitted
07 Aug. 2022
Accepted
30 Aug. 2022
First published
01 Okt. 2022

Analyst, 2022,147, 4980-4985

Ultrasensitive detection of DNA methyltransferase activity: a novel dual-amplification fluorescence technique

Q. Wen, D. Li, G. Huang, H. Xi, H. Pan, L. Zhang, Z. Li, X. Xiao and W. Zhu, Analyst, 2022, 147, 4980 DOI: 10.1039/D2AN01302A

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