Issue 22, 2022

Quantifying DNA damage on paper sensors via controlled template-independent DNA polymerization

Abstract

We report on a paper-based sensor capable of performing template-independent DNA synthesis by terminal deoxynucleotidyl transferase (TdT). Importantly, we observed that TdT efficiently incorporates fluorescently labeled dUTP on to 3′-OH ends of DNA strands in a strictly controllable manner on cellulose paper, in comparison to its distributive mode of DNA synthesis in solution. Due to the high roughness and porous nature of cellulose paper, we attribute this controllable DNA polymerization to the pore confinement effect on the catalytic behaviour of TdT. Taking advantage of this finding, we proposed a paper-assisted TdT (PAT) assay for absolute quantification of alkylated DNA lesions (N7-methylguanine), DNA deamination (cytosine-to-uracil) and DNA oxidation (8-oxo-7,8-dihydroguanine) by combining various DNA glycosylases. This PAT assay provides a low-cost, high throughput and easy to use method for quantifying the absolute levels of various types of DNA lesions, thus making it well-suited for drug development, genotoxicity testing, and environmental toxicology.

Graphical abstract: Quantifying DNA damage on paper sensors via controlled template-independent DNA polymerization

Supplementary files

Article information

Article type
Edge Article
Submitted
04 Aug 2021
Accepted
13 Oct 2021
First published
20 Apr 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2022,13, 6496-6501

Quantifying DNA damage on paper sensors via controlled template-independent DNA polymerization

W. Xue, Q. Zhang, Y. Chang, J. D. Brennan, Y. Li and M. Liu, Chem. Sci., 2022, 13, 6496 DOI: 10.1039/D1SC04268H

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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