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Triple‐Targeting Delivery of CRISPR/Cas9 To Reduce the Risk of Cardiovascular Diseases
Author(s) -
Zhang Lingmin,
Wang Le,
Xie Yangzhouyun,
Wang Peng,
Deng Sai,
Qin Aiping,
Zhang Jiangjiang,
Yu Xiyong,
Zheng Wenfu,
Jiang Xingyu
Publication year - 2019
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201903618
Subject(s) - crispr , nanoclusters , cas9 , nanocarriers , genome editing , chemistry , cancer research , cholesterol , mutagenesis , computational biology , mutation , microbiology and biotechnology , biochemistry , medicine , biology , pharmacology , gene , drug , organic chemistry
A high level of low‐density lipoprotein cholesterol (LDL‐C) in the blood is a major risk factor for coronary heart disease. Herein, we present a triple‐targeting strategy to generate a loss‐of‐function mutation in Pcsk9 , which regulates plasma cholesterol levels, using a nanocarrier‐delivered CRISPR/Cas9 system. Nuclear localization signal (NLS)‐tagged Cas9 and Pcsk9 ‐targeted single guide RNA (sgPcsk9) were complexed with gold nanoclusters (GNCs) modified with cationic HIV‐1‐transactivating transcriptor (TAT) peptide and further encapsulated in a galactose‐modified lipid layer to target the nanoclusters to the liver. The resulting nanoclusters had an in vitro Pcsk9 ‐editing efficiency of about 60 % and resulting in a decrease in plasma LDL‐C in mice of approximately 30%. No off‐target mutagenesis was detected in 10 sites with high similarity. This approach may have therapeutic potential for the prevention and treatment of cardiovascular disease without side effects.