TY - JOUR
T1 - Investigation of genomic DNA methylation by ultraviolet resonant Raman spectroscopy
AU - D'Amico, Francesco
AU - Zucchiatti, Paolo
AU - Latella, Katia
AU - Pachetti, Maria
AU - Gessini, Alessandro
AU - Masciovecchio, Claudio
AU - Vaccari, Lisa
AU - Pascolo, Lorella
N1 - Funding Information:
The authors are grateful to the CERIC‐ERIC for granting the beamtime 20172056. L.P. acknowledges the support from the Institute from Maternal and Child Health, IRCCS Burlo Garofolo (5mille15D1). The authors are grateful to Dr. Martina Bradaschia (IRCSS Burlo Garofolo) for assistance with the manuscript editing.
Publisher Copyright:
© 2020 Wiley-VCH GmbH
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/12
Y1 - 2020/12
N2 - 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.
AB - 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.
KW - cytosine nucleotides
KW - DNA methylation
KW - epigenetics
KW - isolated DNA
KW - UV resonant Raman
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U2 - 10.1002/jbio.202000150
DO - 10.1002/jbio.202000150
M3 - Article
C2 - 32729213
AN - SCOPUS:85091322038
VL - 13
JO - Journal of Biophotonics
JF - Journal of Biophotonics
SN - 1864-063X
IS - 12
M1 - e202000150
ER -