Nanoparticle Delivery of CRISPR/Cas9 for Genome Editing

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.

Functionalized Nanoparticles Targeting Tumor-Associated Macrophages as Cancer Therapy

“…NP-based CRISPR/Cas9 delivery systems represent a novel technology in which NPs instead of viruses are employed to directly deliver the CRISPR/Cas9 complex, or plasmids expressing Cas9 and guide RNAs, to the nuclei of targeted cells. In recent years, many NP formulations have been described as suitable non-viral delivery systems for CRISPR/Cas9, including lipid, polymeric and inorganic NPs [ 274 ]. Several molecules, such as CD47 (“do not eat me” signal), which are implicated in tumor immune escape mechanisms and can be targeted by small-molecule drugs [ 4 ], have now become targets of gene-editing approaches utilizing NPs as a delivery system for CRISPR/Cas9.…”

Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing

“…Like lipid nanoparticles, polymer-based nanoparticles can also transverse the complex in the membrane through endocytosis. 60 Viral vectors are the natural experts for in vivo CRISPR/Cas-9 delivery. 61 Vectors, such as adenoviral vectors (AVs), adeno-associated viruses (AAVs), and lentivirus vectors (LVs) are currently being widely used as delivery methods due to their higher delivery efficiency relative to physical and chemical methods.…”

Polyethyleneimine-Based Lipopolyplexes as Carriers in Anticancer Gene Therapies

“…Recently, there has been an abundance of research focused on novel delivery methods for this versatile and precise genome editing tool, as efficient delivery remains the main obstacle in utilizing it in gene therapies. For the time being, formation of stearyl octaarginine-based lipopolyplexes was proposed [ 124 ], although polyethyleneimine derivatives alone, as well as various liposomal or exosomal formulations, have been used for effective CRISPR/Cas system delivery, and they have been reviewed recently [ 125 , 126 ]. The next step would be now to combine both of those approaches together.…”