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Published January 3, 2018 | Published
Journal Article Open

In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the SVZ


Directed evolution continues to expand the capabilities of complex biomolecules for a range of applications, such as adeno-associated virus vectors for gene therapy; however, advances in library design and selection strategies are key to develop variants that overcome barriers to clinical translation. To address this need, we applied structure-guided SCHEMA recombination of the multimeric adeno-associated virus (AAV) capsid to generate a highly diversified chimeric library with minimal structural disruption. A stringent in vivo Cre-dependent selection strategy was implemented to identify variants that transduce adult neural stem cells (NSCs) in the subventricular zone. A novel variant, SCH9, infected 60% of NSCs and mediated 24-fold higher GFP expression and a 12-fold greater transduction volume than AAV9. SCH9 utilizes both galactose and heparan sulfate as cell surface receptors and exhibits increased resistance to neutralizing antibodies. These results establish the SCHEMA library as a valuable tool for directed evolution and SCH9 as an effective gene delivery vector to investigate subventricular NSCs.

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© 2017 The American Society of Gene and Cell Therapy. Under a Creative Commons license (Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)) Published online: September 08, 2017; Accepted: September 3, 2017; Received: June 18, 2017. D.S.O. was supported by a National Science Foundation Graduate Fellowship and a UC Berkeley Dissertation Fellowship. S.S. was supported by a National Defense Science and Engineering Graduate Fellowship. J.L.S.-O. was supported by a National Science Foundation Graduate Fellowship and a UC Berkeley Graduate Division Fellowship. This work was also supported by NIH grant R01 EY022975. The authors are grateful to Timothy Day and Dr. John Flannery of UC Berkeley for providing pRepHelper and pSub2RepKO, Dr. Jan Carette of Stanford for the AAVR cell line, Dr. Marla Feller of UC Berkeley for the Ai9 tdTomato mice, and Dr. Thomas Gaj for helpful discussions. Conflicts of Interest: D.V.S., D.S.O., and J.L.S.-O. are inventors on patents involving AAVdirected evolution. D.V.S. is also a co-founder of a company developing AAV vectors for clinical gene therapy.

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August 19, 2023
August 19, 2023