posted on 2016-11-13, 00:00authored byLing Li, Linjiang Song, Xiaowei Liu, Xi Yang, Xia Li, Tao He, Ning Wang, Suleixin Yang, Chuan Yu, Tao Yin, Yanzhu Wen, Zhiyao He, Xiawei Wei, Weijun Su, Qinjie Wu, Shaohua Yao, Changyang Gong, Yuquan Wei
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.