Sendai virus, an RNA virus with no risk of genomic integration, delivers CRISPR/Cas9 for efficient gene editing.
Sendai virus, an RNA virus with no risk of genomic integration, delivers CRISPR/Cas9 for efficient gene editing.
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DOI：
10.1038/mtm.2016.57
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Lee B
中科院分区：
文献类型：
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作者：
Park A;Hong P;Won ST;Thibault PA;Vigant F;Oguntuyo KY;Taft JD;Lee B
RNA引导的核酸内切酶（RGEN）介导的基因编辑（特别是通过CRISPR/Cas9）的出现刺激了人们为提高RGEN递送和靶向诱变的效率而进行的大量努力。用于递送Cas9及其相关指导RNA、慢病毒和腺相关病毒系统的主要病毒载体具有不期望的随机整合到宿主基因组中的潜力。在这里，我们重新利用仙台病毒（一种没有病毒DNA相且仅在细胞质中复制的RNA病毒）作为高效Cas9介导的基因编辑的递送系统。仙台病毒感染的高效率导致在不存在任何选择的情况下，细胞系（在各种内源性和转基因座处为75-98%）和原代人单核细胞（在ccr 5基因座处为88%）中的高靶向诱变率。结合仙台病毒作为一种有前途的基因治疗载体的广泛工作，可以感染包括造血干细胞在内的各种细胞类型，这项概念验证研究为使用仙台病毒以及其他相关的副粘病毒作为基因编辑的通用和有效工具打开了大门。
The advent of RNA-guided endonuclease (RGEN)-mediated gene editing, specifically via CRISPR/Cas9, has spurred intensive efforts to improve the efficiency of both RGEN delivery and targeted mutagenesis. The major viral vectors in use for delivery of Cas9 and its associated guide RNA, lentiviral and adeno-associated viral systems, have the potential for undesired random integration into the host genome. Here, we repurpose Sendai virus, an RNA virus with no viral DNA phase and that replicates solely in the cytoplasm, as a delivery system for efficient Cas9-mediated gene editing. The high efficiency of Sendai virus infection resulted in high rates of on-target mutagenesis in cell lines (75–98% at various endogenous and transgenic loci) and primary human monocytes (88% at the ccr5 locus) in the absence of any selection. In conjunction with extensive former work on Sendai virus as a promising gene therapy vector that can infect a wide range of cell types including hematopoietic stem cells, this proof-of-concept study opens the door to using Sendai virus as well as other related paramyxoviruses as versatile and efficient tools for gene editing.