Leverage of Nuclease-deficient CasX for Preventing Pathological Angiogenesis

Overview

Journal Mol Ther Nucleic Acids

Publisher Cell Press

Date 2023 Sep 4

PMID 37662968

Authors

Haote Han

Yanhui Yang

Yunjuan Jiao

Hui Qi

Zhuo Han

Luping Wang

Lijun Dong

Jingkui Tian

Bart Vanhaesebroeck

Xiaopeng Li

Junwen Liu

Gaoen Ma

Hetian Lei

Affiliations

Soon will be listed here.

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%) expression of p110δ, the catalytic subunit of PI3Kδ, encoded by . 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 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.

Citing Articles

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