The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has revolutionized the field of gene editing. Continuous efforts in developing this technology have enabled efficient in vitro, ex vivo, and in vivo gene editing through a variety of delivery strategies. Viral vectors are commonly used in in vitro, ex vivo, and in vivo delivery systems, but they can cause insertional mutagenesis, have limited cloning capacity, and/or elicit immunologic responses. Physical delivery methods are largely restricted to in vitro and ex vivo systems, whereas chemical delivery methods require extensive optimization to improve their efficiency for in vivo gene editing. Achieving a safe and efficient in vivo delivery system for CRISPR/Cas9 remains the most challenging aspect of gene editing. Recently, extracellular vesicle-based systems were reported in various studies to deliver Cas9 in vitro and in vivo. In comparison with other methods, extracellular vesicles offer a safe, transient, and cost-effective yet efficient platform for delivery, indicating their potential for Cas9 delivery in clinical trials. In this review, we first discuss the pros and cons of different Cas9 delivery strategies. We then specifically review the development of extracellular vesicle-mediated gene editing and highlight the strengths and weaknesses of this technology.

中文翻译：

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CRISPR / Cas9递送策略的最新进展。

簇状规则间隔的短回文重复序列（CRISPR）/ Cas9系统彻底改变了基因编辑领域。通过不断地努力开发该技术，可以通过多种递送策略进行有效的体外，离体和体内基因编辑。病毒载体通常用于体外，离体和体内递送系统，但是它们可引起插入诱变，克隆能力有限和/或引起免疫应答。物理递送方法主要限于体外和离体系统，而化学递送方法需要广泛的优化以提高其体内基因编辑的效率。为CRISPR / Cas9实现安全高效的体内递送系统仍然是基因编辑中最具挑战性的方面。最近，在各种研究中报道了基于细胞外囊泡的系统以在体外和体内递送Cas9。与其他方法相比，细胞外囊泡提供了一个安全，短暂且经济高效的高效转运平台，表明它们在临床试验中具有Cas9转运的潜力。在本文中，我们首先讨论了不同的Cas9投放策略的利弊。然后，我们专门回顾了细胞外囊泡介导的基因编辑的发展，并强调了这项技术的优缺点。我们首先讨论不同Cas9投放策略的优缺点。然后，我们专门回顾了细胞外囊泡介导的基因编辑的发展，并强调了该技术的优缺点。我们首先讨论不同Cas9投放策略的优缺点。然后，我们专门回顾了细胞外囊泡介导的基因编辑的发展，并强调了该技术的优缺点。