title: Efficient transduction and optogenetic stimulation of retinal bipolar cells by a synthetic adeno-associated virus capsid and promoter.
abstract: In this report, we describe the development of a modified adeno-associated virus (AAV) capsid and promoter for transduction of retinal ON-bipolar cells. The bipolar cells, which are post-synaptic to the photoreceptors, are important retinal targets for both basic and preclinical research. In particular, a therapeutic strategy under investigation for advanced forms of blindness involves using optogenetic molecules to render ON-bipolar cells light-sensitive. Currently, delivery of adequate levels of gene expression is a limiting step for this approach. The synthetic AAV capsid and promoter described here achieves high level of optogenetic transgene expression in ON-bipolar cells. This evokes high-frequency (~100 Hz) spiking responses in ganglion cells of previously blind, rd1, mice. Our vector is a promising vehicle for further development toward potential clinical use.
authors.affiliation: Center for Advanced Retinal and Ophthalmic Therapeutics, F.M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland jebennet@mail.med.upenn.edu therese.cronin@fmi.ch.
authors.forename: Therese
authors.initials: T
authors.lastname: Cronin
authors.affiliation: Center for Advanced Retinal and Ophthalmic Therapeutics, F.M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA, USA.
authors.forename: Luk H
authors.initials: LH
authors.lastname: Vandenberghe
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Péter
authors.initials: P
authors.lastname: Hantz
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Josephine
authors.initials: J
authors.lastname: Juttner
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Reimann
authors.affiliation: Faculty of Physics, Babes-Bolyai University, Cluj-Napoca, Romania.
authors.forename: Agota-Enikő
authors.initials: AE
authors.lastname: Kacsó
authors.affiliation: Center for Advanced Retinal and Ophthalmic Therapeutics, F.M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Huckfeldt
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland Genetics Department, Harvard Medical School, Boston, MA, USA.
authors.forename: Volker
authors.initials: V
authors.lastname: Busskamp
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Hubertus
authors.initials: H
authors.lastname: Kohler
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Lagali
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Botond
authors.initials: B
authors.lastname: Roska
authors.affiliation: Center for Advanced Retinal and Ophthalmic Therapeutics, F.M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA jebennet@mail.med.upenn.edu therese.cronin@fmi.ch.
authors.forename: Jean
authors.initials: J
authors.lastname: Bennett
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D051245:Retinal Bipolar Cells / Q000821:virology*; D014161:Transduction, Genetic / Q000379:methods*
keywords: adeno‐associated virus; capsid library; multi‐electrode array; optogenetics; promoter optimization
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