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Title

Quantitative analysis of dose dependent DNA fragmentation in dry pBR322 plasmid using long read sequencing and Monte Carlo simulations.

Authors

Beaudier, Pierre; Zein, Sara A.; Chatzipapas, Konstantinos; Ngoc Tran, Hoang; Devès, Guillaume; Plawinski, Laurent; Liénard, Rémy; Dupuy, Denis; Barberet, Philippe; Incerti, Sébastien; Gobet, Franck; Seznec, Hervé

Abstract

Exposure to ionizing radiation can induce genetic aberrations via unrepaired DNA strand breaks. To investigate quantitatively the dose–effect relationship at the molecular level, we irradiated dry pBR322 plasmid DNA with 3 MeV protons and assessed fragmentation yields at different radiation doses using long-read sequencing from Oxford Nanopore Technologies. This technology applied to a reference DNA model revealed dose-dependent fragmentation, as evidenced by read length distributions, showing no discernible radiation sensitivity in specific genetic sequences. In addition, we propose a method for directly measuring the single-strand break (SSB) yield. Furthermore, through a comparative study with a collection of previous works on dry DNA irradiation, we show that the irradiation protocol leads to biases in the definition of ionizing sources. We support this scenario by discussing the size distributions of nanopore sequencing reads in the light of Geant4 and Geant4-DNA simulation toolkit predictions. We show that integrating long-read sequencing technologies with advanced Monte Carlo simulations paves a promising path toward advancing our comprehension and prediction of radiation-induced DNA fragmentation.

Subjects

MONTE Carlo method; DNA damage; RADIATION doses; RADIATION damage; RADIATION exposure

Publication

Scientific Reports, 2024, Vol 14, Issue 1, p1

ISSN

2045-2322

Publication type

Academic Journal

DOI

10.1038/s41598-024-69406-3

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