Multiparametric domain insertional profiling of adeno-associated virus VP1
Year: 2023
Type: article
Abstract: Evolved properties of Adeno-Associated Virus (AAV), such as broad tropism and immunogenicity in humans, are barriers to AAV-based gene therapy. Previous efforts to re-engineer these properties have focused on variable regions near AAV’s 3-fold protrusions and capsid protein termini. To comprehensively survey AAV capsids for engineerable hotspots, we determined multiple AAV fitness phenotypes upon insertion of large, structured protein domains into the entire AAV-DJ capsid protein VP1. This is the largest and most comprehensive AAV domain insertion dataset to date. Our data revealed a surprising robustness of AAV capsids to accommodate large domain insertions. There was strong positional, domain-type, and fitness phenotype dependence of insertion permissibility, which clustered into correlated structural units that we could link to distinct roles in AAV assembly, stability, and infectivity. We also identified new engineerable hotspots of AAV that facilitate the covalent attachment of binding scaffolds, which may represent an alternative approach to re-direct AAV tropism.
Institutions: University of Minnesota, Hormel (United States)
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,900
Funders: National Science Foundation, University of Minnesota, National Institutes of Health, University of Illinois at Chicago, Materials Research Science and Engineering Center, Harvard University, Genomics Center, University of Minnesota, National Institute on Drug Abuse, National Institute of General Medical Sciences, Division of Materials Research, Division of Electrical, Communications and Cyber Systems
Awards: G1801249269, G259601046, G310967942, G403942265, G4470453470, G5316173330, G5449242509, G6917505600, G6995829561, G743970376, G7573957695, University of Minnesota Materials Research Science and Engineering Center, NNCI: Midwest Nano Infrastructure Corridor (MINIC), Decoding mechanotransduction mechanisms of cell-surface receptors, Center for Neural Circuits in Addiction, Viral vector technology for cell type specific gene delivery
OpenAlex ID: W4387956086