{"doi":"10.1002/ijch.202300111","title":"A Quantitative Sequencing Method for 5‐Formylcytosine in RNA","abstract":"Abstract 5‐Formylcytosine (f 5 C) modification is present in human mitochondrial methionine tRNA (mt‐tRNA Met ) and cytosolic leucine tRNA (ct‐tRNA Leu ), with their formation mediated by NSUN3 and ALKBH1. f 5 C has also been detected in yeast mRNA and human tRNA, but its transcriptome‐wide distribution in mammals has not been studied. Here we report f 5 C‐seq, a quantitative sequencing method to map f 5 C transcriptome‐wide in HeLa and mouse embryonic stem cells (mESCs). We show that f 5 C in RNA can be reduced to dihydrouracil (DHU) by pic‐borane, and DHU can be exclusively read as T during reverse transcription (RT) reaction, allowing the detection and quantification of f 5 C sites by a unique C‐to‐T mutation signature. We validated f 5 C‐seq by identifying and quantifying the two known f 5 C sites in tRNA, in which the f 5 C modification fractions dropped significantly in ALKBH1‐depleted cells. By applying f 5 C‐seq to chromatin‐associated RNA (caRNA), we identified several highly modified f 5 C sites in HeLa and mouse embryonic stem cells (mESC).","journal":"Israel Journal of Chemistry","year":2023,"id":359836,"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":10,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.945,"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":622153,"name":"Kinga Pajdzik","orcid":null,"position":1,"is_corresponding":false},{"id":247085,"name":"Hui‐Lung Sun","orcid":"0000-0003-2273-311X","position":2,"is_corresponding":false},{"id":247089,"name":"Linda Zhang","orcid":"0000-0001-9486-419X","position":3,"is_corresponding":false},{"id":1111314,"name":"Li‐Sheng Zhang","orcid":null,"position":4,"is_corresponding":false},{"id":239945,"name":"Tong Wu","orcid":"0000-0001-9968-9909","position":5,"is_corresponding":false},{"id":841798,"name":"Lei Yang","orcid":"0000-0002-3029-8114","position":6,"is_corresponding":false},{"id":371532,"name":"Tao Pan","orcid":"0000-0003-1096-8405","position":7,"is_corresponding":false},{"id":21408,"name":"Chuan He","orcid":"0000-0003-4319-7424","position":8,"is_corresponding":false},{"id":233670,"name":"Qing Dai","orcid":"0000-0002-8578-1568","position":9,"is_corresponding":false},{"id":262498,"name":"Ruitu Lyu","orcid":"0000-0003-4597-1604","position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-19T01:13:57.361401Z","pmid":"40492241","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":[]}