Investigation of genomic DNA methylation by ultraviolet resonant Raman spectroscopy.
DNA methylation
UV resonant Raman
cytosine nucleotides
epigenetics
isolated DNA
Journal
Journal of biophotonics
ISSN: 1864-0648
Titre abrégé: J Biophotonics
Pays: Germany
ID NLM: 101318567
Informations de publication
Date de publication:
12 2020
12 2020
Historique:
received:
21
04
2020
revised:
24
07
2020
accepted:
28
07
2020
pubmed:
31
7
2020
medline:
24
6
2021
entrez:
31
7
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32729213
doi: 10.1002/jbio.202000150
doi:
Substances chimiques
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202000150Informations de copyright
© 2020 Wiley-VCH GmbH.
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