UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Leverage of nuclease-deficient CasX for preventing pathological angiogenesis

Han, H; Yang, Y; Jiao, Y; Qi, H; Han, Z; Wang, L; Dong, L; ... Lei, H; + view all (2023) Leverage of nuclease-deficient CasX for preventing pathological angiogenesis. Molecular Therapy - Nucleic Acids , 33 pp. 738-748. 10.1016/j.omtn.2023.08.001. Green open access

[thumbnail of 1-s2.0-S2162253123002135-main.pdf]
Preview
PDF
1-s2.0-S2162253123002135-main.pdf - Published Version

Download (5MB) | Preview

Abstract

Gene editing with a CRISPR/Cas system is a novel potential strategy for treating human diseases. Pharmacological inhibition of phosphoinositide 3-kinase (PI3K) δ suppresses retinal angiogenesis in a mouse model of oxygen-induced retinopathy. Here we show that an innovative system of adeno-associated virus (AAV)-mediated CRISPR/nuclease-deficient (d)CasX fused with the Krueppel-associated box (KRAB) domain is leveraged to block (81.2% ± 6.5%) in vitro expression of p110δ, the catalytic subunit of PI3Kδ, encoded by Pik3cd. This CRISPR/dCasX-KRAB (4, 269 bp) system is small enough to be fit into a single AAV vector. We then document that recombinant AAV serotype (rAAV)1 efficiently transduces vascular endothelial cells from pathologic retinal vessels, which show high expression of p110δ; furthermore, we demonstrate that blockade of retinal p110δ expression by intravitreally injected rAAV1-CRISPR/dCasX-KRAB targeting the Pik3cd promoter prevents (32.1% ± 5.3%) retinal p110δ expression as well as pathological retinal angiogenesis in a mouse model of oxygen-induced retinopathy. These data establish a strong foundation for treating pathological angiogenesis by AAV-mediated CRISPR interference with p110δ expression.

Type: Article
Title: Leverage of nuclease-deficient CasX for preventing pathological angiogenesis
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.omtn.2023.08.001
Publisher version: https://doi.org/10.1016/j.omtn.2023.08.001
Language: English
Additional information: © 2023 The Author(s). Original content in this paper is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/).
Keywords: MT: RNA/DNA editing, CRISPR/dCasX-KRAB, rAAV1, Pik3cd, PI3Kδ, oxygen-induced retinopathy, angiogenesis
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Oncology
URI: https://discovery.ucl.ac.uk/id/eprint/10175836
Downloads since deposit
11Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item