{"doi":"10.1093/hmg/ddae144","title":"Delivering large genes using adeno-associated virus and the CRE-lox DNA recombination system","abstract":"Adeno-associated virus (AAV) is a safe and efficient gene delivery vehicle for gene therapies. However, its relatively small packaging capacity limits its use as a gene transfer vector. Here, we describe a strategy to deliver large genes that exceed the AAV's packaging capacity using up to four AAV vectors and the CRE-lox DNA recombination system. We devised novel lox sites by combining non-compatible and reaction equilibrium-modifying lox site variants. These lox sites facilitate sequence-specific and near-unidirectional recombination of AAV vector genomes, enabling efficient reconstitution of up to 16 kb of therapeutic genes in a pre-determined configuration. Using this strategy, we have developed AAV gene therapy vectors to deliver IFT140, PCDH15, CEP290, and CDH23 and demonstrate efficient production of full-length proteins in cultured mammalian cells and mouse retinas. Notably, AAV-IFT140 gene therapy vectors ameliorated retinal degeneration and preserved visual functions in an IFT140-associated retinitis pigmentosa mouse model. The CRE-lox approach described here provides a simple, flexible, and effective platform for generating AAV gene therapy vectors beyond AAV's packaging capacity.","journal":"Human Molecular Genetics","year":2024,"id":431785,"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":16,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9531,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":442985,"name":"Kun-Do Rhee","orcid":null,"position":1,"is_corresponding":false},{"id":1236453,"name":"Rylee J Staudt","orcid":null,"position":2,"is_corresponding":false},{"id":904763,"name":"Jacob M. Thompson","orcid":"0009-0007-2960-5889","position":3,"is_corresponding":false},{"id":505973,"name":"Ying Hsu","orcid":"0000-0002-0396-9680","position":4,"is_corresponding":false},{"id":1236043,"name":"Salma Hassan","orcid":"0000-0001-9116-0357","position":5,"is_corresponding":false},{"id":354469,"name":"Arlene V. Drack","orcid":"0000-0002-8195-8426","position":6,"is_corresponding":false},{"id":416384,"name":"Seongjin Seo","orcid":"0000-0002-7908-3210","position":7,"is_corresponding":false},{"id":416383,"name":"Poppy Datta","orcid":"0000-0003-4989-0975","position":0,"is_corresponding":true}],"reference_count":66,"raw_metadata":null,"created_at":"2026-07-19T01:59:30.747263Z","pmid":"39393808","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":[]}