CRISPR delivery with extracellular vesicles: Promises and challenges

Abstract

The CRISPR gene editing tool holds great potential for curing genetic disorders. However, the safe, efficient, and specific delivery of the CRISPR/Cas9 components into cells and tissues remains a challenge. While many currently available delivery methods achieve high levels of gene editing effects in vivo, they often result in genotoxicity and immunogenicity. Extracellular vesicles (EVs), which are cell‐derived lipid nanoparticles, are capable of transferring protein and nucleic acid cargoes between cells, making them a promising endogenous alternative to synthetic delivery methods. This review provides a comprehensive analysis of the currently available strategies for EV‐mediated delivery of CRISPR/Cas9. These strategies include cell‐based, passive loading obtained by overexpression of CRISPR/Cas9, active loading involving protein or RNA dimerization, and loading into already purified EVs. All these approaches suggest that EV‐based CRISPR/Cas9 delivery is useful for achieving both in vitro and in vivo gene editing. Despite that, substantial variations in cellular uptake and gene editing efficiencies indicate that further improvement and standardization are required for the therapeutic use of EVs as a CRISPR/Cas9 delivery vehicle. These improvements include, but is not limited to, the high‐yield purification of EVs, increased loading and release efficiencies, as well as improved tissue‐ or cell‐specific targeting specificities.

Exosomes: A Cutting-Edge Theranostics Tool for Oral Cancer

“…189 Genetically modified exosomes such as exosome-based CRISPR system delivery in cancer therapy are more effective. 190 In an exosome-based cancer therapy developed process, plant sources get lot of attention due to their lack of toxicity and medicinal plants get more priority in future clinical applications. 183 Several in vivo methods (migration assay, angiogenesis analysis, MMPs protein analysis, cell cycle analysis) and in vivo methods (tumor size analysis, immune histochemistry analysis after treatment, skin sensitivity-for toxicological safety analysis) have been examined for effectiveness as therapeutic exosomes.…”

“…CAR-T cell derived exosomes are less toxic compared to CAR-T cell therapy in cancer, and chimeric exosome’s effective anticancer activity is proved in in vitro and in vivo models . Genetically modified exosomes such as exosome-based CRISPR system delivery in cancer therapy are more effective . In an exosome-based cancer therapy developed process, plant sources get lot of attention due to their lack of toxicity and medicinal plants get more priority in future clinical applications .…”

Fusion of Exosomes and Nanotechnology: Cutting-Edge Cancer Theranostics

“…An exosome–liposome hybrid enhances exosomes encapsulation capacity, and this method is used in gene editing . CRISPR gene editing is a promising tool for cancer-therapeutics, but cell and tissue-level transport is a challenging event . Exosomes assist in overcoming this complication and developing an effective gene therapy approach. , Exosomes-based CRISPR delivery reprograms decreases oncogene activity and suppresses tumor growth .…”

Strategies for enhanced gene delivery to the central nervous system

“…However, further improvements are needed to optimize gene editing efficiency and cell specificity. 78 At present, two primary methodologies are employed for loading EVs: direct loading of purified EVs and cell-based loading. In direct loading, EVs are loaded after purification, while in cell-based loading, loading occurs either passively via CRISPR/Cas9 overexpression or actively through interactions involving proteins or RNA.…”