National Medical Research Center of Tuberculosis and Infectious Diseases, Ministry of Health, Moscow 127994, Russia.
Institute of Immunology, Federal Medical Biological Agency, Moscow 115522, Russia.
Int J Mol Sci. 2020 Oct 5;21(19):7362. doi: 10.3390/ijms21197362.
CRISPR/Cas technologies have advanced dramatically in recent years. Many different systems with new properties have been characterized and a plethora of hybrid CRISPR/Cas systems able to modify the epigenome, regulate transcription, and correct mutations in DNA and RNA have been devised. However, practical application of CRISPR/Cas systems is severely limited by the lack of effective delivery tools. In this review, recent advances in developing vehicles for the delivery of CRISPR/Cas in the form of ribonucleoprotein complexes are outlined. Most importantly, we emphasize the use of extracellular vesicles (EVs) for CRISPR/Cas delivery and describe their unique properties: biocompatibility, safety, capacity for rational design, and ability to cross biological barriers. Available molecular tools that enable loading of desired protein and/or RNA cargo into the vesicles in a controllable manner and shape the surface of EVs for targeted delivery into specific tissues (e.g., using targeting ligands, peptides, or nanobodies) are discussed. Opportunities for both endogenous (intracellular production of CRISPR/Cas) and exogenous (post-production) loading of EVs are presented.
CRISPR/Cas 技术近年来取得了显著进展。已经对许多具有新特性的不同系统进行了描述，并设计了大量能够修饰表观基因组、调节转录以及纠正 DNA 和 RNA 突变的杂交 CRISPR/Cas 系统。然而，CRISPR/Cas 系统的实际应用受到有效传递工具缺乏的严重限制。在这篇综述中，概述了以核糖核蛋白复合物形式递送 CRISPR/Cas 的载体的最新进展。最重要的是，我们强调了使用细胞外囊泡（EVs）进行 CRISPR/Cas 递送，并描述了它们独特的特性：生物相容性、安全性、合理设计的能力以及跨越生物屏障的能力。讨论了可用于以可控方式将所需蛋白质和/或 RNA 货物装入囊泡并为靶向递送至特定组织（例如，使用靶向配体、肽或纳米抗体）塑造 EV 表面的可用分子工具。介绍了内源性（细胞内产生 CRISPR/Cas）和外源性（生产后）装载 EV 的机会。
