{"doi":"10.1101/2023.01.27.525935","title":"pA regulator: a system for controlling mammalian gene expression via the modulation of polyA signal cleavage","abstract":"Abstract The ability to control the expression of a therapeutic gene or a transgene in mammalian cells is crucial for safe and efficacious gene and cell therapy, as well as for elucidating the function of a specific gene product. Yet current mammalian gene regulation systems either evoke harmful immune responses in hosts or lack the required regulatory efficiency. Here we describe a highly responsive RNA-based molecular switch, the pA regulator, that harnesses the power of polyA signal cleavage within the 5’ UTR to control mammalian gene expression. The pA regulator is governed by a ‘dual mechanism’ to ensure maximal control of gene expression: (1) aptamer clamping of polyA signal via drug binding and (2) drug-induced alternative splicing that removes the polyA signal. The metholology achieves an induction efficiency up to 900-fold with an EC 50 of 0.5μg/ml Tetracycline, a drug concentration that falls well within the FDA-approved dose range. The pA regulator circumvents the immune responses that plague other systems by eliminating the use of a regulatory foreign protein and the need to change transgene coding sequences. Furthermore, it is not dependent on any specific promoter, therefore the system is simple to implement in a single non-viral or viral vector. In a mouse study using AAV-mediated gene transfer, we showed that the pA regulator controlled transgene expression in a “dose-dependent’ and “reversible” manner and exhibited long-term stability in vivo , in which both features are crucial for effective therapeutics. The pA regulator is the first non-immunogenic system that demonstrates an EC 50 at a drug concentration approved by FDA, making it a clinically relevant gene regulation system that could open a new window of opportunity in clinical applications as well as biological studies.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":395724,"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":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9548,"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":1170932,"name":"Jocelyn Duen-Ya Jea","orcid":"0000-0003-3264-2529","position":1,"is_corresponding":false},{"id":3711,"name":"Yan Wang","orcid":"0000-0003-2563-255X","position":2,"is_corresponding":false},{"id":1171285,"name":"Pei-Wen Chao","orcid":null,"position":3,"is_corresponding":false},{"id":1171286,"name":"Laising Yen","orcid":null,"position":4,"is_corresponding":false},{"id":1170931,"name":"Liming Luo","orcid":"0000-0001-8630-5814","position":0,"is_corresponding":true}],"reference_count":47,"raw_metadata":null,"created_at":"2026-07-19T01:19:22.857556Z","pmid":null,"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":[]}