{"doi":"10.17615/p7rb-bh63","title":"rAAV-compatible MiniPromoters for restricted expression in the brain and eye","abstract":"Abstract Background Small promoters that recapitulate endogenous gene expression patterns are important for basic, preclinical, and now clinical research. Recently, there has been a promising revival of gene therapy for diseases with unmet therapeutic needs. To date, most gene therapies have used viral-based ubiquitous promoters–however, promoters that restrict expression to target cells will minimize off-target side effects, broaden the palette of deliverable therapeutics, and thereby improve safety and efficacy. Here, we take steps towards filling the need for such promoters by developing a high-throughput pipeline that goes from genome-based bioinformatic design to rapid testing in vivo. Methods For much of this work, therapeutically interesting Pleiades MiniPromoters (MiniPs; ~4 kb human DNA regulatory elements), previously tested in knock-in mice, were “cut down” to ~2.5 kb and tested in recombinant adeno-associated virus (rAAV), the virus of choice for gene therapy of the central nervous system. To evaluate our methods, we generated 29 experimental rAAV2/9 viruses carrying 19 different MiniPs, which were injected intravenously into neonatal mice to allow broad unbiased distribution, and characterized in neural tissues by X-gal immunohistochemistry for icre, or immunofluorescent detection of GFP. Results The data showed that 16 of the 19 (84 %) MiniPs recapitulated the expression pattern of their design source. This included expression of: Ple67 in brain raphe nuclei; Ple155 in Purkinje cells of the cerebellum, and retinal bipolar ON cells; Ple261 in endothelial cells of brain blood vessels; and Ple264 in retinal Müller glia. Conclusions Overall, the methodology and MiniPs presented here represent important advances for basic and preclinical research, and may enable a paradigm shift in gene therapy.","journal":"UNC Libraries","year":2020,"id":143206,"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":0.9498,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":611273,"name":"Lisa Borretta","orcid":null,"position":1,"is_corresponding":false},{"id":610847,"name":"Michelle X. Zhou","orcid":"0000-0002-0802-5806","position":2,"is_corresponding":false},{"id":611274,"name":"Stéphanie Laprise","orcid":null,"position":3,"is_corresponding":false},{"id":4055,"name":"Robert A. Holt","orcid":"0000-0002-7259-1247","position":4,"is_corresponding":false},{"id":611275,"name":"George Yang","orcid":null,"position":5,"is_corresponding":false},{"id":610848,"name":"Tess C. Lengyell","orcid":"0000-0001-6292-0724","position":6,"is_corresponding":false},{"id":610849,"name":"Charles N. de Leeuw","orcid":"0000-0001-9413-2095","position":7,"is_corresponding":false},{"id":77770,"name":"Wyeth W. Wasserman","orcid":"0000-0001-6098-6412","position":8,"is_corresponding":false},{"id":611276,"name":"Magdalena I. Swanson-Newman","orcid":null,"position":9,"is_corresponding":false},{"id":611277,"name":"Vikramjit Chopra","orcid":null,"position":10,"is_corresponding":false},{"id":611278,"name":"Cletus A. D’Souza","orcid":null,"position":11,"is_corresponding":false},{"id":610850,"name":"Élodie Portales-Casamar","orcid":"0000-0002-9998-7858","position":12,"is_corresponding":false},{"id":265976,"name":"Aravind Asokan","orcid":"0000-0001-5563-4877","position":13,"is_corresponding":false},{"id":611279,"name":"Olga Kaspieva","orcid":null,"position":14,"is_corresponding":false},{"id":611280,"name":"Alice Chou","orcid":null,"position":15,"is_corresponding":false},{"id":610851,"name":"Andrea J. Korecki","orcid":"0000-0001-6532-0695","position":16,"is_corresponding":false},{"id":611281,"name":"Kaelan Wong","orcid":null,"position":17,"is_corresponding":false},{"id":501888,"name":"Garrett E. Berry","orcid":null,"position":18,"is_corresponding":false},{"id":14377,"name":"Steven J.M. Jones","orcid":"0000-0003-3394-2208","position":19,"is_corresponding":false},{"id":610852,"name":"Jack W. Hickmott","orcid":"0000-0003-3764-1138","position":20,"is_corresponding":false},{"id":610853,"name":"Russell J. Bonaguro","orcid":"0000-0003-2581-6252","position":21,"is_corresponding":false},{"id":611282,"name":"Siu Ling Lam","orcid":null,"position":22,"is_corresponding":false},{"id":610854,"name":"Elizabeth M. Simpson","orcid":"0000-0002-0654-4303","position":23,"is_corresponding":false},{"id":610855,"name":"Simone McInerny","orcid":"0000-0002-5048-1549","position":24,"is_corresponding":false},{"id":77640,"name":"Dan Goldowitz","orcid":"0000-0003-4756-4017","position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":null,"created_at":"2026-07-18T23:17:38.432803Z","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":[]}