Chemo-Enzymatic Fluorescence Labeling Of Genomic DNA For Simultaneous Detection Of Global 5-Methylcytosine And 5-Hydroxymethylcytosine.
5-hydroxymethylcytosine
DNA methylation
DNA methyltransferase
epigenetic biomarker
fluorescence
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
17 10 2023
17 10 2023
Historique:
revised:
05
07
2023
received:
29
05
2023
medline:
23
10
2023
pubmed:
31
7
2023
entrez:
30
7
2023
Statut:
ppublish
Résumé
5-Methylcytosine and 5-hydroxymethylcytosine are epigenetic modifications involved in gene regulation and cancer. We present a new, simple, and high-throughput platform for multi-color epigenetic analysis. The novelty of our approach is the ability to multiplex methylation and de-methylation signals in the same assay. We utilize an engineered methyltransferase enzyme that recognizes and labels all unmodified CpG sites with a fluorescent cofactor. In combination with the already established labeling of the de-methylation mark 5-hydroxymethylcytosine via enzymatic glycosylation, we obtained a robust platform for simultaneous epigenetic analysis of these marks. We assessed the global epigenetic levels in multiple samples of colorectal cancer and observed a 3.5-fold reduction in 5hmC levels but no change in DNA methylation levels between sick and healthy individuals. We also measured epigenetic modifications in chronic lymphocytic leukemia and observed a decrease in both modification levels (5-hydroxymethylcytosine: whole blood 30 %; peripheral blood mononuclear cells (PBMCs) 40 %. 5-methylcytosine: whole blood 53 %; PBMCs 48 %). Our findings propose using a simple blood test as a viable method for analysis, simplifying sample handling in diagnostics. Importantly, our results highlight the assay's potential for epigenetic evaluation of clinical samples, benefiting research and patient management.
Identifiants
pubmed: 37518671
doi: 10.1002/cbic.202300400
doi:
Substances chimiques
5-hydroxymethylcytosine
1123-95-1
5-Methylcytosine
6R795CQT4H
DNA
9007-49-2
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202300400Subventions
Organisme : European Research Council
ID : 817811
Pays : International
Organisme : NHGRI NIH HHS
ID : R01HG009190
Pays : United States
Informations de copyright
© 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.
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