title: In Silico Reconstruction of the Viral Evolutionary Lineage Yields a Potent Gene Therapy Vector.
abstract: Adeno-associated virus (AAV) vectors have emerged as a gene-delivery platform with demonstrated safety and efficacy in a handful of clinical trials for monogenic disorders. However, limitations of the current generation vectors often prevent broader application of AAV gene therapy. Efforts to engineer AAV vectors have been hampered by a limited understanding of the structure-function relationship of the complex multimeric icosahedral architecture of the particle. To develop additional reagents pertinent to further our insight into AAVs, we inferred evolutionary intermediates of the viral capsid using ancestral sequence reconstruction. In-silico-derived sequences were synthesized de novo and characterized for biological properties relevant to clinical applications. This effort led to the generation of nine functional putative ancestral AAVs and the identification of Anc80, the predicted ancestor of the widely studied AAV serotypes 1, 2, 8, and 9, as a highly potent in vivo gene therapy vector for targeting liver, muscle, and retina.
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Zinn
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Pacouret
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Vadim
authors.initials: V
authors.lastname: Khaychuk
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Heikki T
authors.initials: HT
authors.lastname: Turunen
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Livia S
authors.initials: LS
authors.lastname: Carvalho
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Andres-Mateos
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Samiksha
authors.initials: S
authors.lastname: Shah
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Rajani
authors.initials: R
authors.lastname: Shelke
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Anna C
authors.initials: AC
authors.lastname: Maurer
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Plovie
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA.
authors.forename: Ru
authors.initials: R
authors.lastname: Xiao
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA. Electronic address: luk_vandenberghe@meei.harvard.edu.
authors.forename: Luk H
authors.initials: LH
authors.lastname: Vandenberghe
mesh_terms: D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data