Prime editing brings immense promise to correct a large number of human pathogenic mutations and enact diverse edit types without introducing widespread undesired editing events. Delivery of prime editors in vivo would enable such edits to be introduced in a clinical setting. The coding sequence for prime editor, however, is too large to fit within the size-constrained adeno-associated virus (AAV) genome. Herein, we describe a split Staphylococcus aureus prime editor capable of being delivered by dual AAVs. We characterize the editing ability of plasmid-based versions of an S. aureus prime editor in vitro at a variety of loci with diverse edit types. We investigate various split prime editor architectures and alternative dimerization domains. Finally, we demonstrate the capacity of prime editor to be co-delivered by dual AAVs in vitro. While editing rates are lower than desired, this approach presents an important step to translate prime editing for in vivo delivery.

中文翻译：

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分离的金黄色葡萄球菌用于AAV递送的主要编辑器

优质的编辑带来了巨大的希望，可以纠正大量的人类致病突变并制定各种编辑类型，而不会引起广泛的不良编辑事件。在体内交付主要编辑人员将使此类编辑可以在临床环境中引入。但是，主要编辑器的编码序列太大，无法放入受大小限制的腺相关病毒（AAV）基因组中。在这里，我们描述了一个分裂的金黄色葡萄球菌主要编辑器，能够通过双重AAV传递。我们表征了基于质粒的金黄色葡萄球菌主要编辑器在体外在具有不同编辑类型的多个基因座上的编辑能力。我们研究了各种拆分的主要编辑器体系结构和替代的二聚化域。最后，我们展示了主编辑器可以通过双重AAV在体外共同交付的能力。