Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice
Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice
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DOI：
10.1021/acsnano.6b04261
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发表时间：
2017-01-01
期刊：
影响因子：
17.1
通讯作者：
Wei, Yuquan
中科院分区：
文献类型：
--
作者：
Li, Ling;Song, Linjiang;Wei, Yuquan
CRISPR-Cas9已经成为一个多功能的基因组编辑平台。然而，由于常用的CRISPR-Cas9系统尺寸较大，其有效递送一直是一个挑战，并限制了其在基础研究和治疗应用中的效用。本文中，构建了多功能核靶向“核-壳”人工病毒（RRPHC）用于递送CRISPR-Cas9系统。人工病毒可以有效地负载CRISPR-Cas9系统，加速内体逃逸，并在没有额外的核定位信号的情况下促进渗透到细胞核中，从而实现靶向基因破坏。值得注意的是，人工病毒比SuperFect、Lipofectamine 2000和Lipofectamine 3000更有效。当装载CRISPR-Cas9质粒时，它诱导比Lipofectamine 3000更高的靶向基因破坏功效。此外，人工病毒通过双受体介导的内吞作用有效地靶向卵巢癌，并且副作用最小。当装载有靶向MTH 1基因的Cas9-hMTH 1系统时，RRPHC显示出在体内对MTH 1的有效破坏。这种策略可以适用于在体内递送CRISPR-Cas9质粒或其他功能性核酸。
CRISPR-Cas9 has emerged as a versatile genome-editing platform. However, due to the large size of the commonly used CRISPR-Cas9 system, its effective delivery has been a challenge and limits its utility for basic research and therapeutic applications. Herein, a multifunctional nucleus-targeting "core-shell" artificial virus (RRPHC) was constructed for the delivery of CRISPR-Cas9 system. The artificial virus could efficiently load with the CRISPR-Cas9 system, accelerate the endosomal escape, and promote the penetration into the nucleus without additional nuclear-localization signal, thus enabling targeted gene disruption. Notably, the artificial virus is more efficient than SuperFect, Lipofectamine 2000, and Lipofectamine 3000. When loaded with a CRISPR-Cas9 plasmid, it induced higher targeted gene disruption efficacy than that of Lipofectamine 3000. Furthermore, the artificial virus effectively targets the ovarian cancer via dual-receptor mediated endocytosis and had minimum side effects. When loaded with the Cas9-hMTH1 system targeting MTH1 gene, RRPHC showed effective disruption of MTH1 in vivo. This strategy could be adapted for delivering CRISPR-Cas9 plasmid or other functional nucleic acids in vivo.