Year: 2018
Type: article
Abstract: 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.
Source: Molecular Therapy
Institutions: University of California San Diego, Guangzhou Women and Children Medical Center, Guangzhou Medical University, Salk Institute for Biological Studies, The University of Sydney, Children's Medical Research Institute, Wojskowy Instytut Higieny i Epidemiologii, Duke University, VA San Diego Healthcare System
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
Funders: U.S. Department of Veterans Affairs, Burroughs Wellcome Fund, March of Dimes Foundation, Consejo Nacional de Ciencia y Tecnología, University of California, San Diego, National Human Genome Research Institute, National Cancer Institute, National Institute of General Medical Sciences, Richard P. Kimmel and Laurine Kimmel Charitable Foundation
OpenAlex ID: W2800887634