{"doi":"10.1016/j.omtm.2020.03.007","title":"Site-Directed Mutagenesis Improves the Transduction Efficiency of Capsid Library-Derived Recombinant AAV Vectors","abstract":"Recombinant adeno-associated virus (rAAV) vectors selected from capsid libraries present enormous advantages in high selectivity of tissue tropism and their potential use in human gene therapy applications. For example, rAAV-LK03, was used in a gene therapy trial for hemophilia A (ClinicalTrials.gov: NCT03003533). However, high doses in patients resulted in severe adverse events and subsequent loss of factor VIII (FVIII) expression. Thus, additional strategies are needed to enhance the transduction efficiency of capsid library-derived rAAV vectors such that improved clinical efficacy can be achieved at low vector doses. In this study, we characterized two commonly used library-derived rAAV vectors, rAAV-DJ and rAAV-LK03. It was concluded that rAAV-DJ shared similar transport pathways (e.g., cell surface binding, endocytosis-dependent internalization, and cytoplasmic trafficking) with rAAV serotype 2, while rAAV-LK03 and rAAV serotype 3 shared similar transport pathways. We then performed site-directed mutagenesis of surface-exposed tyrosine (Y), serine (S), aspartic acid (D), and tryptophan (W) residues on rAAV-DJ and rAAV-LK03 capsids. Our results demonstrated that rAAV-DJ-S269T and rAAV-LK03-Y705+731F variants had significantly enhanced transduction efficiency compared to wild-type counterparts. Our studies suggest that the strategy of site-directed mutagenesis should be applicable to other non-natural AAV variants for their optimal use in human gene therapy.","journal":"Molecular Therapy — Methods & Clinical Development","year":2020,"id":100558,"datarank":1.1174599344094434,"base_score":3.367295829986474,"endowment":3.367295829986474,"self_citation_contribution":0.5050943744979712,"citation_network_contribution":0.612365559911472,"self_endowment_contribution":0.5050943744979712,"citer_contribution":0.612365559911472,"corpus_percentile":null,"corpus_rank":null,"citation_count":28,"citer_count":22,"citers_with_citation_signal":19,"citers_with_endowment":19,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9492,"is_data_producer":true,"deposit_databanks":{"ClinicalTrials.gov":["NCT03003533"]},"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":493635,"name":"X Chen","orcid":"0000-0001-6155-6035","position":1,"is_corresponding":false},{"id":493636,"name":"Yilin Xie","orcid":"0000-0003-1452-0800","position":2,"is_corresponding":false},{"id":494660,"name":"Qingyun Zheng","orcid":null,"position":3,"is_corresponding":false},{"id":493637,"name":"Jinyan Xie","orcid":"0000-0002-5406-4195","position":4,"is_corresponding":false},{"id":494661,"name":"Chenghui Yu","orcid":null,"position":5,"is_corresponding":false},{"id":493638,"name":"Nikéa Pittman","orcid":"0000-0002-1629-2488","position":6,"is_corresponding":false},{"id":493639,"name":"Sixian Qi","orcid":"0000-0002-6831-0289","position":7,"is_corresponding":false},{"id":493640,"name":"Fa‐Xing Yu","orcid":"0000-0003-4834-9335","position":8,"is_corresponding":false},{"id":311796,"name":"Mavis Agbandje‐McKenna","orcid":"0000-0003-0565-5578","position":9,"is_corresponding":false},{"id":385394,"name":"Arun Srivastava","orcid":"0000-0003-1168-8034","position":10,"is_corresponding":false},{"id":415528,"name":"Chen Ling","orcid":"0000-0003-1527-1471","position":11,"is_corresponding":false},{"id":415523,"name":"Gai Ran","orcid":"0000-0003-0900-1715","position":0,"is_corresponding":true}],"reference_count":38,"raw_metadata":null,"created_at":"2026-07-18T22:39:01.863288Z","pmid":"32258217","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":[]}