biostudies-literatureZinn ENEI NIH HHS100803https://www.ebi.ac.uk/biostudies/studies/S-EPMC9729830biostudies-literatureUnknown3(11)Gene therapy is emerging as a modality in 21st-century medicine. Adeno-associated viral (AAV) gene transfer is a leading technology to achieve efficient and durable expression of a therapeutic transgene. However, the structural complexity of the capsid has constrained efforts to engineer the particle toward improved clinical safety and efficacy. Here, we generate a curated library of barcoded AAVs with mutations across a variety of functionally relevant motifs. We then screen this library in vitro and in vivo in mice and nonhuman primates, enabling a broad, multiparametric assessment of every vector within the library. Among the results, we note a single residue that modulates liver transduction across all interrogated models while preserving transduction in heart and skeletal muscles. MorCell reports. MedicineAncestral library identifies conserved reprogrammable liver motif on AAV capsid.PMC9729830P30 EY003790Unzu CGurrala RSanmiguel JVandenberghe LHWagers AJMessemer KSantos JAndres-Mateos ETurunen HTDiop CTipper CBhatt UFieldsend AMerkel ERios CZabaleta NPeacker BZinn EEstelien RSchmit PFfalseAncestral library identifies conserved reprogrammable liver motif on AAV capsid.Gene therapy is emerging as a modality in 21st-century medicine. Adeno-associated viral (AAV) gene transfer is a leading technology to achieve efficient and durable expression of a therapeutic transgene. However, the structural complexity of the capsid has constrained efforts to engineer the particle toward improved clinical safety and efficacy. Here, we generate a curated library of barcoded AAVs with mutations across a variety of functionally relevant motifs. We then screen this library in vitro and in vivo in mice and nonhuman primates, enabling a broad, multiparametric assessment of every vector within the library. Among the results, we note a single residue that modulates liver transduction across all interrogated models while preserving transduction in heart and skeletal muscles. Mor2022-01-01T00:00:00Z2022 Nov2026-05-28T02:58:01.028Z2025-04-19T12:11:18.914ZS-EPMC97298303632797310.1016/j.xcrm.2022.100803