Investigation of genomic DNA methylation by ultraviolet resonant Raman spectroscopy

Francesco D'Amico, Paolo Zucchiatti, Katia Latella, Maria Pachetti, Alessandro Gessini, Claudio Masciovecchio, Lisa Vaccari, Lorella Pascolo

Research output: Contribution to journalArticlepeer-review

Abstract

Cytosine plays a preeminent role in DNA methylation, an epigenetic mechanism that regulates gene expression, the misregulation of which can lead to severe diseases. Several methods are nowadays employed for assessing the global DNA methylation levels, but none of them combines simplicity, high sensitivity, and low operating costs to be translated into clinical applications. Ultraviolet (UV) resonant Raman measurements at excitation wavelengths of 272 nm, 260 nm, 250 nm, and 228 nm have been carried out on isolated deoxynucleoside triphosphates (dNTPs), on a dNTP mixture as well as on genomic DNA (gDNA) samples, commercial from salmon sperm and non-commercial from B16 murine melanoma cell line. The 228 nm excitation wavelength was identified as the most suitable energy for enhancing cytosine signals over the other DNA bases. The UV Raman measurements performed at this excitation wavelength on hyper-methylated and hypo-methylated DNA from Jurkat leukemic T-cell line have revealed significant spectral differences with respect to gDNA isolated from salmon sperm and mouse melanoma B16 cells. This demonstrates how the proper choice of the excitation wavelength, combined with optimized extraction protocols, makes UV Raman spectroscopy a suitable technique for highlighting the chemical modifications undergone by cytosine nucleotides in gDNA upon hyper- and hypo-methylation events.

Original languageEnglish
Article numbere202000150
JournalJournal of Biophotonics
Volume13
Issue number12
DOIs
Publication statusPublished - Dec 2020

Keywords

  • cytosine nucleotides
  • DNA methylation
  • epigenetics
  • isolated DNA
  • UV resonant Raman

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Engineering(all)
  • Physics and Astronomy(all)

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