{"doi":"10.1101/2025.10.15.682674","title":"Splice-switching antisense oligonucleotides correct cryptic exon inclusion and restore SDCCAG8 protein in Bardet-Biedl Syndrome","abstract":"ABSTRACT Bardet-Biedl Syndrome (BBS) is a ciliopathy often associated with progressive blindness and obesity. A patient presenting with BBS was discovered to have two mutations within 55bp of each other in intron 7 of SDCCAG8 ( BBS16 ). One of the biallelic mutations, c.740+356C>T, causes inclusion of cryptic exon(s) containing premature termination codons, while c.740+301G>A has not been characterized. We hypothesized that antisense oligonucleotides (ASOs) complementary to the patient’s mutations or to the cryptic exon splice sites would correct the splicing of SDCCAG8 between exons 7 and 8 to prevent cryptic exon inclusion and restore SDCCAG8 expression. We systematically screened 20nt-long ASOs tiled across each mutation and ASOs targeting the 3′ splice sites of the cryptic exons in patient-derived fibroblasts, using RT-PCR assays to assess exon 7 and 8 splicing. We identified one ASO for each mutation and a cryptic exon-targeting ASO that restored the splicing pattern to that observed in an unaffected cell line. Lead ASOs were further investigated through RT-PCR, RNA sequencing, and western blotting to confirm ASO-mediated restoration of wild-type transcript and protein. Notably, ASO 20, which targets the cryptic exon 7a/7a′ splice site rather than patient-specific mutations, achieved the greatest rescue effect, increasing exon 7-8 splicing from 0% to an average of 26% and restoring SDCCAG8 protein from undetectable levels to approximately 40% of wild-type expression. This mutation-agnostic approach could benefit multiple patients with cryptic exon inclusion in this region of SDCCAG8 , expanding therapeutic impact beyond traditional N-of-1 ASO strategies. These findings establish a molecular foundation for clinical development of ASO therapy for BBS caused by SDCCAG8 splicing defects.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":578147,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9599,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":831992,"name":"Bailey L McCurdy","orcid":"0000-0002-1073-2673","position":1,"is_corresponding":false},{"id":301131,"name":"Austin Larson","orcid":"0000-0002-2079-6929","position":2,"is_corresponding":false},{"id":280111,"name":"Emily A. McCourt","orcid":null,"position":3,"is_corresponding":false},{"id":694194,"name":"Michael L. Kaufman","orcid":"0000-0003-2441-5836","position":4,"is_corresponding":false},{"id":294718,"name":"Amy E. Campbell","orcid":"0000-0001-6513-5836","position":5,"is_corresponding":false},{"id":447397,"name":"Chad G. Pearson","orcid":"0000-0003-1915-6593","position":6,"is_corresponding":false},{"id":632157,"name":"Scott Demarest","orcid":"0000-0002-4544-3724","position":7,"is_corresponding":false},{"id":262478,"name":"J. Matthew Taliaferro","orcid":"0000-0001-7580-1433","position":8,"is_corresponding":false},{"id":87044,"name":"Jay R. Hesselberth","orcid":"0000-0002-6299-179X","position":9,"is_corresponding":false},{"id":667918,"name":"Sujatha Jagannathan","orcid":"0000-0001-9039-2631","position":10,"is_corresponding":false},{"id":1488195,"name":"Kelleen E McEntee","orcid":null,"position":0,"is_corresponding":true}],"reference_count":20,"raw_metadata":null,"created_at":"2026-07-19T02:58:20.638044Z","pmid":"41279107","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":[]}