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

Ojala, David S. and Sun, Sabrina and Santiago-Ortiz, Jorge L. and Shapiro, Mikhail G. and Romero, Philip A. and Schaffer, David V. (2018) In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the SVZ. Molecular Therapy, 26 (1). pp. 304-319. ISSN 1525-0016. http://resolver.caltech.edu/CaltechAUTHORS:20180215-134720732

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Abstract

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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.ymthe.2017.09.006 DOIArticle
https://www.sciencedirect.com/science/article/pii/S1525001617304185PublisherArticle
ORCID:
AuthorORCID
Shapiro, Mikhail G.0000-0002-0291-4215
Additional Information:© 2017 The American Society of Gene and Cell Therapy. 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.
Funders:
Funding AgencyGrant Number
NSF Graduate Research FellowshipUNSPECIFIED
University of California, BerkeleyUNSPECIFIED
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
NIHR01 EY022975
Subject Keywords:gene therapy, adeno-associated virus, capsid engineering, neural stem cell, subventricular zone, directed evolution, CNS
Record Number:CaltechAUTHORS:20180215-134720732
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20180215-134720732
Official Citation:David S. Ojala, Sabrina Sun, Jorge L. Santiago-Ortiz, Mikhail G. Shapiro, Philip A. Romero, David V. Schaffer, In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the SVZ, Molecular Therapy, Volume 26, Issue 1, 2018, Pages 304-319, ISSN 1525-0016, https://doi.org/10.1016/j.ymthe.2017.09.006. (http://www.sciencedirect.com/science/article/pii/S1525001617304185)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84851
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Feb 2018 22:53
Last Modified:21 Feb 2018 17:06

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