Abstract:
Efficient genome editing by using CRISPR technologies requires simultaneous and efficient delivery of multiple genetically encoded components to mammalian cells. Amongst all editing approaches, prime editing (PE) has the unique potential to perform seamless genome rewriting, in the absence of DNA double-strand breaks (DSBs). The cargo capacity required for efficient PE delivery to mammalian cells stands at odd with the limited packaging capacity of traditional viral delivery vectors. By contrast, baculovirus (BV) has a large synthetic DNA capacity and can efficiently transduce mammalian cells. Here we describe a protocol for the assembly of baculovirus vectors for multiplexed prime editing in mammalian cells.
摘要:
通过使用CRISPR技术进行有效的基因组编辑需要将多种遗传编码的组分同时有效地递送到哺乳动物细胞。在所有编辑方法中,素编辑(PE)具有执行无缝基因组重写的独特潜力,在没有DNA双链断裂(DSB)的情况下。有效的PE递送至哺乳动物细胞所需的货物容量与传统病毒递送载体的有限包装容量不同。相比之下,杆状病毒(BV)具有很大的合成DNA容量,可以有效地转导哺乳动物细胞。在这里,我们描述了用于哺乳动物细胞中多重引物编辑的杆状病毒载体组装方案。