title: An evolved adeno-associated viral variant enhances gene delivery and gene targeting in neural stem cells.
abstract: Gene delivery to, and gene targeting in, stem cells would be a highly enabling technology for basic science and biomedical application. Adeno-associated viral (AAV) vectors have demonstrated the capacity for efficient delivery to numerous cells, but their application to stem cells has been limited by low transduction efficiency. Due to their considerable advantages, however, engineering AAV delivery systems to enhance gene delivery to stem cells may have an impact in stem cell biology and therapy. Therefore, using several diverse AAV capsid libraries-including randomly mutagenized, DNA shuffled, and random peptide insertion variants-we applied directed evolution to create a "designer" AAV vector with enhanced delivery efficiency for neural stem cells (NSCs). A novel AAV variant, carrying an insertion of a selected peptide sequence on the surface of the threefold spike within the heparin-binding site, emerged from this evolution. Importantly, this evolved AAV variant mediated efficient gene delivery to rat, mouse, and human NSCs, as well as efficient gene targeting within adult NSCs, and it is thus promising for applications ranging from basic stem cell biology to clinical translation.
authors.affiliation: Department of Chemical Biomolecular Engineering, University of California, Berkeley, California, USA.
authors.forename: Jae-Hyung
authors.initials: JH
authors.lastname: Jang
authors.forename: James T
authors.initials: JT
authors.lastname: Koerber
authors.forename: Jung-Suk
authors.initials: JS
authors.lastname: Kim
authors.forename: Prashanth
authors.initials: P
authors.lastname: Asuri
authors.forename: Tandis
authors.initials: T
authors.lastname: Vazin
authors.forename: Melissa
authors.initials: M
authors.lastname: Bartel
authors.forename: Albert
authors.initials: A
authors.lastname: Keung
authors.forename: Inchan
authors.initials: I
authors.lastname: Kwon
authors.forename: Kook In
authors.initials: KI
authors.lastname: Park
authors.forename: David V
authors.initials: DV
authors.lastname: Schaffer
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002845:Chromatography; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D058953:Neural Stem Cells / Q000821:virology*; D016133:Polymerase Chain Reaction; D051381:Rats; D011916:Rats, Inbred F344