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MADDD-seq, a novel massively parallel sequencing tool for simultaneous detection of DNA damage and mutations

Affiliations
  • 1University of Southern California, Leonard Davis School of Gerontology, Los Angeles, CA, USA.
  • 2Translational Research Program, Public Health Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
  • 3Data Core, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
  • 4Department of Biological Sciences, Mississippi State University, Mississippi State, MS, USA.
  • 5Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, USA.

Published on:

August 16, 2024

Abstract

Our genome is exposed to a wide variety of DNA-damaging agents. If left unrepaired, this damage can be converted into mutations that promote carcinogenesis or the development of genetically inherited diseases. As a result, researchers and clinicians require tools that can detect DNA damage and mutations with exceptional sensitivity. In this study, we describe a massively parallel sequencing tool termed Mutation And DNA Damage Detection-seq (MADDD-seq) that is capable of detecting O6-methyl guanine lesions and mutations simultaneously, with a single assay. To illustrate the dual capabilities of MADDD-seq, we treated WT and DNA repair deficient yeast cells with the DNA-damaging agent MNNG and tracked DNA lesions and mutations over a 24-h time period. This approach allowed us to identify thousands of DNA adducts and mutations in a single sequencing run and gain deep insight into the kinetics of DNA repair and mutagenesis.