TY - JOUR
T1 - Engineered Extracellular Vesicle-Delivered CRISPR/CasRx as a Novel RNA Editing Tool
AU - Li, Tianwen
AU - Zhang, Liansheng
AU - Lu, Tao
AU - Zhu, Tongming
AU - Feng, Canbin
AU - Gao, Ni
AU - Liu, Fei
AU - Yu, Jingyu
AU - Chen, Kezhu
AU - Zhong, Junjie
AU - Tang, Qisheng
AU - Zhang, Quan
AU - Deng, Xiangyang
AU - Ren, Junwei
AU - Zeng, Jun
AU - Zhou, Haibo
AU - Zhu, Jianhong
N1 - Publisher Copyright:
© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.
PY - 2023/4/5
Y1 - 2023/4/5
N2 - Engineered extracellular vesicles (EVs) are considered excellent delivery vehicles for a variety of therapeutic agents, including nucleic acids, proteins, drugs, and nanomaterials. Recently, several studies have indicated that clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) delivered by EVs enable efficient DNA editing. However, an RNA editing tool delivered by EVs is still unavailable. Here, a signal peptide-optimized and EVs-delivered guide RNA (gRNA) and CRISPR/CasRx (Cas13d) system capable of rapidly inhibiting the expression of targeted genes with quick catabolism after performing their functions is developed. EVs with CRISPR/CasRx and tandem gRNAs targeting pivotal cytokines are further packed whose levels increase substantially over the course of acute inflammatory diseases and find that these engineered EVs inhibit macrophage activation in vitro. More importantly, this system attenuates lipopolysaccharide (LPS)-triggered acute lung injury and sepsis in the acute phase, mitigating organ damage and improving the prognosis in vivo. In summary, a potent tool is provided for short-acting RNA editing, which could be a powerful therapeutic platform for the treatment of acute diseases.
AB - Engineered extracellular vesicles (EVs) are considered excellent delivery vehicles for a variety of therapeutic agents, including nucleic acids, proteins, drugs, and nanomaterials. Recently, several studies have indicated that clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) delivered by EVs enable efficient DNA editing. However, an RNA editing tool delivered by EVs is still unavailable. Here, a signal peptide-optimized and EVs-delivered guide RNA (gRNA) and CRISPR/CasRx (Cas13d) system capable of rapidly inhibiting the expression of targeted genes with quick catabolism after performing their functions is developed. EVs with CRISPR/CasRx and tandem gRNAs targeting pivotal cytokines are further packed whose levels increase substantially over the course of acute inflammatory diseases and find that these engineered EVs inhibit macrophage activation in vitro. More importantly, this system attenuates lipopolysaccharide (LPS)-triggered acute lung injury and sepsis in the acute phase, mitigating organ damage and improving the prognosis in vivo. In summary, a potent tool is provided for short-acting RNA editing, which could be a powerful therapeutic platform for the treatment of acute diseases.
KW - CRISPR/CasRx
KW - RNA editing
KW - extracellular vesicles
KW - inflammatory disease
UR - http://www.scopus.com/inward/record.url?scp=85147441689&partnerID=8YFLogxK
U2 - 10.1002/advs.202206517
DO - 10.1002/advs.202206517
M3 - Article
AN - SCOPUS:85147441689
SN - 2198-3844
VL - 10
JO - Advanced Science
JF - Advanced Science
IS - 10
M1 - 2206517
ER -