PDF(Pubmed)
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.
摘要：
CRISPR基因编辑工具在治疗遗传疾病方面具有巨大潜力。然而,保险箱,高效,以及将CRISPR/Cas9组分特异性递送到细胞和组织中仍然是一个挑战。虽然许多目前可用的递送方法在体内实现了高水平的基因编辑效应，它们通常导致遗传毒性和免疫原性。细胞外囊泡（EV），它们是细胞衍生的脂质纳米颗粒，能够在细胞之间转移蛋白质和核酸物质，使它们成为合成递送方法的有希望的内源性替代品。这篇综述全面分析了目前可用的EV介导的CRISPR/Cas9递送策略。这些策略包括基于细胞的，通过过表达CRISPR/Cas9获得的被动加载，涉及蛋白质或RNA二聚化的主动加载，并装载到已经纯化的电动汽车中。所有这些方法表明基于EV的CRISPR/Cas9递送对于实现体外和体内基因编辑都是有用的。尽管如此,细胞摄取和基因编辑效率的显著差异表明,EVs作为CRISPR/Cas9递送载体的治疗用途需要进一步改进和标准化.这些改进包括,但不限于,电动汽车的高产量纯化，增加装载和释放效率，以及改善的组织或细胞特异性靶向特异性。
