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Abstract：
Clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology has revolutionized the field of gene therapy as it has enabled precise genome editing with unprecedented accuracy and efficiency, paving the way for clinical applications to treat otherwise incurable genetic disorders. Typically, precise genome editing requires the delivery of multiple components to the target cells that, depending on the editing platform used, may include messenger RNA (mRNA), protein complexes, and DNA fragments. For clinical purposes, these have to be efficiently delivered into transplantable cells, such as primary T lymphocytes or hematopoietic stem and progenitor cells that are typically sensitive to exogenous substances. This challenge has limited the broad applicability of precise gene therapy applications to those strategies for which efficient delivery methods are available. Electroporation-based methodologies have been generally applied for gene editing applications, but procedure-associated toxicity has represented a major burden. With the advent of novel and less disruptive methodologies to deliver genetic cargo to transplantable cells, it is now possible to safely and efficiently deliver multiple components for precise genome editing, thus expanding the applicability of these strategies. In this review, we describe the different delivery systems available for genome editing components, including viral and non-viral systems, highlighting their advantages, limitations, and recent clinical applications. Recent improvements to these delivery methods to achieve cell specificity represent a critical development that may enable in vivo targeting in the future and will certainly play a pivotal role in the gene therapy field.
摘要：
聚集的定期穿插短回文重复序列（CRISPR）/CRISPR相关蛋白9（Cas9）技术彻底改变了基因治疗领域，因为它以前所未有的准确性和效率实现了精确的基因组编辑。为治疗无法治愈的遗传疾病的临床应用铺平了道路。通常,精确的基因组编辑需要将多种成分传递给靶细胞，根据使用的编辑平台，可能包括信使RNA(mRNA)，蛋白质复合物，和DNA片段。为了临床目的，这些必须有效地传递到可移植的细胞中，例如通常对外源物质敏感的原代T淋巴细胞或造血干细胞和祖细胞。这种挑战已将精确基因治疗应用的广泛适用性限制在可获得有效递送方法的那些策略上。基于电穿孔的方法已普遍应用于基因编辑应用，但与程序相关的毒性是一个主要负担。随着新型和破坏性较小的方法的出现，将遗传货物运送到可移植的细胞，现在可以安全有效地提供多种成分进行精确的基因组编辑，从而扩大了这些策略的适用性。在这次审查中,我们描述了可用于基因组编辑组件的不同递送系统，包括病毒和非病毒系统，突出他们的优势，局限性,和最近的临床应用。最近对这些递送方法的改进以实现细胞特异性代表了一个关键的发展，该发展可能在将来实现体内靶向，并且肯定会在基因治疗领域发挥关键作用。
