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Nanoparticle Delivery of CRISPR/Cas9 for Genome Editing

Duan, Li, Ouyang, Kan, Xu, Xiao, Xu, Limei, Wen, Caining, Zhou, Xiaoying, Qin, Zhuan, Xu, Zhiyi, Sun, Wei, Liang, Yujie·2021·Frontiers in Genetics
DOI 10.3389/fgene.2021.673286PMC PMC8149999
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Abstract

The emerging clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated system (Cas) gene-editing system represents a promising tool for genome manipulation. However, its low intracellular delivery efficiency severely compromises its use and potency for clinical applications. Nanocarriers, such as liposomes, polymers, and inorganic nanoparticles, have shown great potential for gene delivery. The remarkable development of nanoparticles as non-viral carriers for the delivery of the CRISPR/Cas9 system has shown great promise for therapeutic applications. In this review, we briefly summarize the delivery components of the CRISPR/Cas9 system and report on the progress of nano-system development for CRISPR/Cas9 delivery. We also compare the advantages of various nano-delivery systems and their applications to deliver CRISPR/Cas9 for disease treatment. Nano-delivery systems can be modified to fulfill the tasks of targeting cells or tissues. We primarily emphasize the novel exosome-based CRISPR/Cas9 delivery system. Overall, we review the challenges, development trends, and application prospects of nanoparticle-based technology for CRISPR/Cas9 delivery.

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Crossref · 10.3389/fgene.2021.673286

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Figures

Figure 1 Representative genome editing by three forms of clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9. Cas9 plasmid DNA, RNA, or protein delivery via nanoparticles to be used for precise genome editing. PAM, protospacer adjacent motif.
Figure 1 Representative genome editing by three forms of clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9. Cas9 plasmid DNA, RNA, or protein delivery via nanoparticles to be used for precise genome editing. PAM, protospacer adjacent motif.CC BY · PubMed Central
Figure 2 Rational designs of nano-delivery vehicles for plasmid-, RNA-, and RNP-based CRISPR/Cas9. DNA nanostructures show the controllable size and architecture assembly. Gold nanoparticles show high delivery efficiency.
Figure 2 Rational designs of nano-delivery vehicles for plasmid-, RNA-, and RNP-based CRISPR/Cas9. DNA nanostructures show the controllable size and architecture assembly. Gold nanoparticles show high delivery efficiency.CC BY · PubMed Central
Figure 3 Various engineered exosomes for the efficient delivery of CRISPR/Cas9 components. The co-expression system of SaCas9-GFP1-10 and VSVG-GFP11 can be actively loaded into exosomes. Rapamycin induces an interaction between FK506 binding protein (FKBP)-rapamycin binding domain (FRB) and FKBP, which incorporates the Cas9 complex into the engineered exosome. CD63, as the exosomal member, is fused with GFP and Cas9 protein-linked nanobodies. Thus, the Cas9 protein could be encapsulated into exosomes specifically by the GFP-binding nanobody. CD9-HuR exosomes can actively load AU-enriched mRNA for Cas9 mRNA delivery. Hybrid exosomes with liposomes can effectively load the Cas9 plasmid.
Figure 3 Various engineered exosomes for the efficient delivery of CRISPR/Cas9 components. The co-expression system of SaCas9-GFP1-10 and VSVG-GFP11 can be actively loaded into exosomes. Rapamycin induces an interaction between FK506 binding protein (FKBP)-rapamycin binding domain (FRB) and FKBP, which incorporates the Cas9 complex into the engineered exosome. CD63, as the exosomal member, is fused with GFP and Cas9 protein-linked nanobodies. Thus, the Cas9 protein could be encapsulated into exosomes specifically by the GFP-binding nanobody. CD9-HuR exosomes can actively load AU-enriched mRNA for Cas9 mRNA delivery. Hybrid exosomes with liposomes can effectively load the Cas9 plasmid.CC BY · PubMed Central

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Citations

Cites

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Metadata

Authors
Duan, L., Ouyang, K., Xu, X., Xu, L., Wen, C., Zhou, X., Qin, Z., Xu, Z., Sun, W., & Liang, Y.
Venue
Frontiers in Genetics
Publisher
Frontiers Media SA
Published
2021-05-12
Volume
12
ISSN
1664-8021
DOI
10.3389/fgene.2021.673286
PMC
PMC8149999
Cited by
377
References
14
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
  • National Natural Science Foundation of China (81772394, 81972116)
  • China Postdoctoral Science Foundation (2020M672829)

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