{"doi":"10.1083/jcb.202101092","title":"Catalytically inactive, purified RNase H1: A specific and sensitive probe for RNA–DNA hybrid imaging","abstract":"R-loops are three-stranded nucleic acid structures with both physiological and pathological roles in cells. R-loop imaging generally relies on detection of the RNA-DNA hybrid component of these structures using the S9.6 antibody. We show that the use of this antibody for imaging can be problematic because it readily binds to double-stranded RNA (dsRNA) in vitro and in vivo, giving rise to nonspecific signal. In contrast, purified, catalytically inactive human RNase H1 tagged with GFP (GFP-dRNH1) is a more specific reagent for imaging RNA-DNA hybrids. GFP-dRNH1 binds strongly to RNA-DNA hybrids but not to dsRNA oligonucleotides in fixed human cells and is not susceptible to binding endogenous RNA. Furthermore, we demonstrate that purified GFP-dRNH1 can be applied to fixed cells to detect hybrids after their induction, thereby bypassing the need for cell line engineering. GFP-dRNH1 therefore promises to be a versatile tool for imaging and quantifying RNA-DNA hybrids under a wide range of conditions.","journal":"The Journal of Cell Biology","year":2021,"id":151301,"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":84,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9536,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":575923,"name":"Joshua R. Brickner","orcid":"0000-0002-4620-205X","position":1,"is_corresponding":false},{"id":642958,"name":"Chenlin Song","orcid":"0009-0001-1526-8582","position":2,"is_corresponding":false},{"id":643816,"name":"Su Mon Thin Zar","orcid":null,"position":3,"is_corresponding":false},{"id":330460,"name":"Su S. Maw","orcid":null,"position":4,"is_corresponding":false},{"id":247517,"name":"Frédéric Chédin","orcid":"0000-0002-1306-5335","position":5,"is_corresponding":false},{"id":484856,"name":"Miaw-Sheue Tsai","orcid":"0000-0003-3239-5724","position":6,"is_corresponding":false},{"id":92045,"name":"Karlene A. Cimprich","orcid":"0000-0002-1937-2969","position":7,"is_corresponding":false},{"id":282444,"name":"Magdalena P. Crossley","orcid":"0000-0002-8509-4645","position":0,"is_corresponding":true}],"reference_count":44,"raw_metadata":null,"created_at":"2026-07-18T23:43:11.293086Z","pmid":"34232287","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":[]}