{"doi":"10.3389/fcell.2022.888279","title":"Improved Methods for Deamination-Based m6A Detection","abstract":"N 6 -methyladenosine (m 6 A) is a critical regulator of gene expression and cellular function. Much of our knowledge of m 6 A has been enabled by the identification of m 6 A sites transcriptome-wide. However, global m 6 A profiling methods require high amounts of input RNA to accurately identify methylated RNAs, making m 6 A profiling from rare cell types or scarce tissue samples infeasible. To overcome this issue, we previously developed DART-seq, which relies on the expression of a fusion protein consisting of the APOBEC1 cytidine deaminase tethered to the m 6 A-binding YTH domain. APOBEC1-YTH directs C-to-U mutations adjacent to m 6 A sites, therefore enabling single nucleotide-resolution m 6 A mapping. Here, we present an improved version of DART-seq which utilizes a variant of the YTH domain engineered to achieve enhanced m 6 A recognition. In addition, we develop in vitro DART-seq and show that it performs similarly to cellular DART-seq and can map m 6 A in any sample of interest using nanogram amounts of total RNA. Altogether, these improvements to the DART-seq approach will enable better m 6 A detection and will facilitate the mapping of m 6 A in samples not previously amenable to global m 6 A profiling.","journal":"Frontiers in Cell and Developmental Biology","year":2022,"id":265192,"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":11,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9562,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":924297,"name":"Xinhe Yin","orcid":null,"position":1,"is_corresponding":false},{"id":274676,"name":"Christopher L. Holley","orcid":"0000-0002-2870-3352","position":2,"is_corresponding":false},{"id":318745,"name":"Kate D. Meyer","orcid":"0000-0001-7197-4054","position":3,"is_corresponding":false},{"id":923780,"name":"Huanyu Zhu","orcid":"0000-0002-1966-9949","position":0,"is_corresponding":true}],"reference_count":39,"raw_metadata":null,"created_at":"2026-07-19T00:26:45.905331Z","pmid":"35573664","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":[]}