In Situ Gene Therapy via AAV-CRISPR-Cas9-Mediated Targeted Gene Regulation

Copyright © 2018 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

Bibliographic Details
Published in:Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 26, No. 7 (2018), p. 1818-1827
Main Author: Moreno, Ana M
Other Involved Persons: Fu, Xin ; Zhu, Jie ; Katrekar, Dhruva ; Shih, Yu-Ru V ; Marlett, John ; Cabotaje, Jessica ; Tat, Jasmine ; Naughton, John ; Lisowski, Leszek ; Varghese, Shyni ; Zhang, Kang ; Mali, Prashant
Format: electronic Article
Item Description:Date Completed 26.06.2019
Date Revised 05.07.2019
published: Print-Electronic
Citation Status MEDLINE
Copyright: From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Physical Description:Online-Ressource
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  • Copyright © 2018 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.
  • Development of efficacious in vivo delivery platforms for CRISPR-Cas9-based epigenome engineering will be critical to enable the ability to target human diseases without permanent modification of the genome. Toward this, we utilized split-Cas9 systems to develop a modular adeno-associated viral (AAV) vector platform for CRISPR-Cas9 delivery to enable the full spectrum of targeted in situ gene regulation functionalities, demonstrating robust transcriptional repression (up to 80%) and activation (up to 6-fold) of target genes in cell culture and mice. We also applied our platform for targeted in vivo gene-repression-mediated gene therapy for retinitis pigmentosa. Specifically, we engineered targeted repression of Nrl, a master regulator of rod photoreceptor determination, and demonstrated Nrl knockdown mediates in situ reprogramming of rod cells into cone-like cells that are resistant to retinitis pigmentosa-specific mutations, with concomitant prevention of secondary cone loss. Furthermore, we benchmarked our results from Nrl knockdown with those from in vivo Nrl knockout via gene editing. Taken together, our AAV-CRISPR-Cas9 platform for in vivo epigenome engineering enables a robust approach to target disease in a genomically scarless and potentially reversible manner