{"doi":"10.1101/2023.08.21.554109","title":"Deep screening of proximal and distal splicing-regulatory elements in a native sequence context","abstract":"ABSTRACT Pre-mRNA splicing, a key process in gene expression, can be therapeutically modulated using various drug modalities, including antisense oligonucleotides (ASOs). However, determining promising targets is impeded by the challenge of systematically mapping splicing-regulatory elements (SREs) in their native sequence context. Here, we use the catalytically dead CRISPR- Rfx Cas13d RNA-targeting system (dCas13d/gRNA) as a programmable platform to bind SREs and modulate splicing by competing against endogenous splicing factors. SpliceRUSH, a high-throughput screening method, was developed to map SREs in any gene of interest using a lentivirus gRNA library that tiles the genetic region, including distal intronic sequences. When applied to SMN2 , a therapeutic target for spinal muscular atrophy, SpliceRUSH robustly identified not only known SREs, but also a novel distal intronic splicing enhancer, which can be targeted to alter exon 7 splicing using either dCas13d/gRNA or ASOs. This technology enables a deeper understanding of splicing regulation with applications for RNA-based drug discovery.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":397104,"datarank":0.16479184330021646,"base_score":1.0986122886681096,"endowment":1.0986122886681096,"self_citation_contribution":0.16479184330021646,"citation_network_contribution":0.0,"self_endowment_contribution":0.16479184330021646,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9545,"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":1004180,"name":"Dmytro Ustianenko","orcid":null,"position":1,"is_corresponding":false},{"id":909174,"name":"Yow-Tyng Yeh","orcid":"0000-0002-3865-8102","position":2,"is_corresponding":false},{"id":1151296,"name":"Xiaojian Wang","orcid":"0000-0001-8232-0558","position":3,"is_corresponding":false},{"id":1000051,"name":"Martin Jacko","orcid":"0000-0001-7378-7439","position":4,"is_corresponding":false},{"id":1154430,"name":"Lekha V. Yesantharao","orcid":"0000-0003-3651-716X","position":5,"is_corresponding":false},{"id":1172719,"name":"Qiyang Wu","orcid":"0000-0002-6230-6000","position":6,"is_corresponding":false},{"id":406994,"name":"Chaolin Zhang","orcid":"0000-0002-8310-7537","position":7,"is_corresponding":false},{"id":291012,"name":"Yocelyn Recinos","orcid":"0000-0002-7377-8660","position":0,"is_corresponding":true}],"reference_count":71,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:19:35.497854Z","pmid":"37662340","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":[]}