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.

Citation： Li Ling, Song Linjiang, Liu Xiaowei, Yang Xi, Li Xia, He Tao, Wang Ning, Yang Suleixin, Yu Chuan, Yin Tao, Wen Yanzhu, He Zhiyao, Wei Xiawei, Su Weijun, Wu Qinjie, Yao Shaohua, Gong Changyang, Wei Yuquan. Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice. West China medical Virtual Journal, 2000, 1(1): 95-111-. doi: 10.1021/acsnano.6b04261 Copy