{"doi":"10.1002/cbic.202300400","title":"Chemo‐Enzymatic Fluorescence Labeling Of Genomic DNA For Simultaneous Detection Of Global 5‐Methylcytosine And 5‐Hydroxymethylcytosine**","abstract":"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.","journal":"ChemBioChem","year":2023,"id":340020,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":16,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9583,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":885474,"name":"Leonie Schütz","orcid":null,"position":1,"is_corresponding":false},{"id":1074012,"name":"Larissa Käver","orcid":"0009-0002-4217-1951","position":2,"is_corresponding":false},{"id":1074339,"name":"Andreas Dankers","orcid":null,"position":3,"is_corresponding":false},{"id":733106,"name":"Sapir Margalit","orcid":null,"position":4,"is_corresponding":false},{"id":733107,"name":"Yael Michaeli","orcid":null,"position":5,"is_corresponding":false},{"id":1074340,"name":"Shahar Zirkin","orcid":null,"position":6,"is_corresponding":false},{"id":809245,"name":"Dmitry Torchinsky","orcid":"0000-0002-9568-6580","position":7,"is_corresponding":false},{"id":732332,"name":"Noa Gilat","orcid":"0000-0001-8185-1605","position":8,"is_corresponding":false},{"id":1074341,"name":"Omer Bahr","orcid":null,"position":9,"is_corresponding":false},{"id":987857,"name":"Gil Nifker","orcid":null,"position":10,"is_corresponding":false},{"id":1074013,"name":"Maya Koren‐Michowitz","orcid":"0000-0002-7223-328X","position":11,"is_corresponding":false},{"id":809247,"name":"Elmar G. Weinhold","orcid":"0000-0002-6481-2257","position":12,"is_corresponding":false},{"id":732334,"name":"Yuval Ebenstein","orcid":"0000-0002-7107-7529","position":13,"is_corresponding":false},{"id":809760,"name":"Sigal Avraham","orcid":null,"position":0,"is_corresponding":true}],"reference_count":51,"raw_metadata":null,"created_at":"2026-07-19T01:10:49.535464Z","pmid":"37518671","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}