title: Artificial Virus Delivers CRISPR-Cas9 System for Genome Editing of Cells in Mice.
abstract: 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.
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Linjiang
authors.initials: L
authors.lastname: Song
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Xi
authors.initials: X
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: He
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Suleixin
authors.initials: S
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yin
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Yanzhu
authors.initials: Y
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Zhiyao
authors.initials: Z
authors.lastname: He
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Xiawei
authors.initials: X
authors.lastname: Wei
authors.affiliation: School of Medicine, Nankai University , Tianjin 300071, P. R. China.
authors.forename: Weijun
authors.initials: W
authors.lastname: Su
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Qinjie
authors.initials: Q
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Changyang
authors.identifier: 0000-0002-2913-0891
authors.initials: C
authors.lastname: Gong
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University , Chengdu 610041, P. R. China.
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D045643:DNA Repair Enzymes / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry; D065546:Drug Liberation; D000072669:Gene Editing / Q000379:methods*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D064593:Heterografts; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics; D010957:Plasmids; D011092:Polyethylene Glycols / Q000737:chemistry*; D014780:Viruses / Q000737:chemistry*
chemical_list: D004337:Drug Carriers; D011092:Polyethylene Glycols; D006820:Hyaluronic Acid; D010744:Phosphoric Monoester Hydrolases; C078207:8-oxodGTPase; D045643:DNA Repair Enzymes
keywords: CRISPR-Cas9; MTH1; artificial virus; genome editing; nucleus-targeting