Designing a single-molecule biophysics tool for characterising DNA damage for techniques that kill infectious pathogens through DNA damage effects

Miller, H; Wollman, AJM; Leake, MC

doi:10.1007/978-3-319-32189-9_9

Designing a single-molecule biophysics tool for characterising DNA damage for techniques that kill infectious pathogens through DNA damage effects

Lookup NU author(s): Dr Adam Wollman ORCiD

Downloads

Full text for this publication is not currently held within this repository. Alternative links are provided below where available.

Abstract

© Springer International Publishing Switzerland 2016. Antibiotics such as the quinolones and fluoroquinolones kill bacterial pathogens ultimately through DNA damage. They target the essential type IIA topoisomerases in bacteria by stabilising the normally transient double-strand break state which is created to modify the supercoiling state of the DNA. Here we discuss the development of these antibiotics and their method of action. Existing methods for DNA damage visualisation, such as the comet assay and immunofluorescence imaging can often only be analysed qualitatively and this analysis is subjective. We describe a putative single-molecule fluorescence technique for quantifying DNA damage via the total fluorescence intensity of a DNA origami tile fully saturated with an intercalating dye, along with the optical requirements for how to implement these into a light microscopy imaging system capable of single-molecule millisecond timescale imaging. This system promises significant improvements in reproducibility of the quantification of DNA damage over traditional techniques.