Stochastic Packaging of Cas Proteins into Exosomes

IF 1.5 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biology Pub Date : 2024-02-01 DOI:10.1134/s0026893324010102

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引用次数: 0

Abstract—CRISPR/Cas systems are perspective molecular tools for targeted manipulation with genetic materials, such as gene editing, regulation of gene transcription, modification of epigenome etc. While CRISPR/Cas systems proved to be highly effective for correcting genetic disorders and treating infectious diseases and cancers in experimental settings, clinical translation of these results is hampered by the lack of efficient CRISPR/Cas delivery vehicles. Modern synthetic nanovehicles based on organic and inorganic polymers have many disadvantages, including toxicity issues, the lack of targeted delivery, and complex and expensive production pipelines. In turn, exosomes are secreted biological nanoparticles that exhibit high biocompatibility, physico-chemical stability, and the ability to cross biological barriers. Early clinical trials found no toxicity associated with exosome injections. In the recent years, exosomes have been considered as perspective delivery vehicles for CRISPR/Cas systems in vivo. The aim of this study was to analyze the efficacy of CRISPR/Cas stochastic packaging into exosomes for several human cell lines. Here, we show that Cas9 protein is effectively localized into the compartment of intracellular exosome biogenesis, but stochastic packaging of Cas9 into exosomes turns to be very low (~1%). As such, stochastic packaging of Cas9 protein is very ineffective and cannot be used for gene editing purposes. Developing novel tools and technologies for loading CRISPR/Cas systems into exosomes is needed.

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将 Cas 蛋白随机包装到外泌体中

摘要--CRISPR/Cas 系统是对遗传物质进行定向操作的分子工具，如基因编辑、基因转录调控、表观基因组修饰等。虽然 CRISPR/Cas 系统在实验环境中被证明在纠正遗传疾病、治疗传染性疾病和癌症方面非常有效，但由于缺乏高效的 CRISPR/Cas 运送载体，这些成果的临床转化受到了阻碍。基于有机和无机聚合物的现代合成纳米载体有许多缺点，包括毒性问题、缺乏靶向递送以及复杂昂贵的生产管道。而外泌体是一种分泌型生物纳米颗粒，具有很高的生物相容性、物理化学稳定性和穿越生物屏障的能力。早期的临床试验发现，注射外泌体没有任何毒性。近年来，外泌体被认为是 CRISPR/Cas 系统在体内的透视传递载体。本研究旨在分析将CRISPR/Cas随机包装到外泌体中对几种人类细胞系的功效。我们在这里发现，Cas9 蛋白能有效地定位到细胞内外泌体生物发生区，但将 Cas9 随机包装到外泌体中的比例非常低（约为 1%）。因此，Cas9 蛋白的随机包装效果很差，不能用于基因编辑目的。因此需要开发新的工具和技术，将CRISPR/Cas系统装入外泌体。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Biology 生物-生化与分子生物学

CiteScore

1.30

自引率

8.30%

发文量

审稿时长

3 months

期刊介绍： Molecular Biology is an international peer reviewed journal that covers a wide scope of problems in molecular, cell and computational biology including genomics, proteomics, bioinformatics, molecular virology and immunology, molecular development biology, molecular evolution and related areals. Molecular Biology publishes reviews, experimental and theoretical works. Every year, the journal publishes special issues devoted to most rapidly developing branches of physical-chemical biology and to the most outstanding scientists.