Cited by 319Extracellular vesicles in diseaseRNA Interference and Gene DeliveryNanoparticle-Based Drug Delivery

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In Vitro and in Vivo RNA Inhibition by CD9-HuR Functionalized Exosomes Encapsulated with miRNA or CRISPR/dCas9

Li, Zhelong, Zhou, Xueying, Wei, Mengying, Gao, Xiaotong, Zhao, Lianbi, Shi, Ruijing, Sun, Wenqi, Duan, Yunyou, Yang, Guodong, Yuan, Lijun·2019·Nano Letters

DOI 10.1021/acs.nanolett.8b02689

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Abstract

In vitro and in vivo delivery of RNAs of interest holds promise for gene therapy. Recently, exosomes are considered as a kind of rational vehicle for RNA delivery, especially miRNA and/or siRNA, while the loading efficiency is limited. In this study, we engineered the exosomes for RNA loading by constructing a fusion protein in which the exosomal membrane protein CD9 was fused with RNA binding protein, while the RNA of interest either natively harbors or is engineered to have the elements for the binding. By proof-of-principle experiments, we here fused CD9 with HuR, an RNA binding protein interacting with miR-155 with a relatively high affinity. In the exosome packaging cells, the fused CD9-HuR successfully enriched miR-155 into exosomes when miR-155 was excessively expressed. Moreover, miR-155 encapsulated in the exosomes in turn could be efficiently delivered into the recipient cells and recognized the endogenous targets. In addition, we also revealed that the CD9-HuR exosomes could enrich the functional miRNA inhibitor or CRISPR/dCas9 when the RNAs were engineered to have the AU rich elements. Taken together, we here have established a novel strategy for enhanced RNA cargo encapsulation into engineered exosomes, which in turn functions in the recipient cells.

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Authors: Li, Z., Zhou, X., Wei, M., Gao, X., Zhao, L., Shi, R., Sun, W., Duan, Y., Yang, G., & Yuan, L.
Affiliations: Tang Du Hospital (CN)
Air Force Medical University (CN)
Corresponding author: Yang, Guodong
Yuan, Lijun
ORCID: Li, Zhelong
Zhou, Xueying
Wei, Mengying
Gao, Xiaotong
Sun, Wenqi
Yang, Guodong
Yuan, Lijun
Venue: Nano Letters
Publisher: American Chemical Society (ACS)
Published: 2018-12-05
Volume: 19
Issue: 1
Pages: 19-28
ISSN: 1530-6984 (ISSN-L)
1530-6992
DOI: 10.1021/acs.nanolett.8b02689
OpenAlex: W2903480993
Cited by: 318
References: 34
Citations by year: 2026: 27 · 2025: 33 · 2024: 54 · 2023: 56 · 2022: 55 · 2021: 43 · 2020: 36 · 2019: 13
FWCI: 8.18
Type: journal article
Language: en
Open access: closed
Locations: Nano Letters (publishedVersion)
PubMed (publishedVersion)
Funders: National Natural Science Foundation of China (81671690, 81871357, 31771507)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords: engineered exosomes, RNA delivery, microRNA delivery, CRISPR/dCas9, miR-155, AU-rich elements, RNA-binding proteins, gene therapy vectors
MeSH terms: ELAV-Like Protein 1, Animals, Cell Line, Humans, RNA-Binding Proteins, Gene Transfer Techniques, RNA, Small Interfering, MicroRNAs, Mice, Exosomes, Tetraspanin 29, CRISPR-Cas Systems

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In Vitro and in Vivo RNA Inhibition by CD9-HuR Functionalized Exosomes Encapsulated with miRNA or CRISPR/dCas9

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