Supramolecular Nanosubstrate-Mediated Delivery for CRISPR/Cas9 Gene Disruption and Deletion

Qian Ban, Peng Yang, Shih Jie Chou, Li Qiao, Haidong Xia, Jingjing Xue, Fang Wang, Xiaobin Xu, Na Sun, Ryan Y. Zhang, Ceng Zhang, Athena Lee, Wenfei Liu, Ting Yi Lin, Yu Ling Ko, Petar Antovski, Xinyue Zhang, Shih Hwa Chiou, Chin Fa Lee, Wenqiao HuiDahai Liu, Steven J. Jonas, Paul S. Weiss, Hsian Rong Tseng*


研究成果: Article同行評審

7 引文 斯高帕斯(Scopus)


The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) is an efficient and precise gene-editing technology that offers a versatile solution for establishing treatments directed at genetic diseases. Currently, CRISPR/Cas9 delivery into cells relies primarily on viral vectors, which suffer from limitations in packaging capacity and safety concerns. These issues with a nonviral delivery strategy are addressed, where Cas9•sgRNA ribonucleoprotein (RNP) complexes can be encapsulated into supramolecular nanoparticles (SMNP) to form RNP⊂SMNPs, which can then be delivered into targeted cells via supramolecular nanosubstrate-mediated delivery. Utilizing the U87 glioblastoma cell line as a model system, a variety of parameters for cellular-uptake of the RNP-laden nanoparticles are examined. Dose- and time-dependent CRISPR/Cas9-mediated gene disruption is further examined in a green fluorescent protein (GFP)-expressing U87 cell line (GFP-U87). The utility of an optimized SMNP formulation in co-delivering Cas9 protein and two sgRNAs that target deletion of exons 45–55 (708 kb) of the dystrophin gene is demonstrated. Mutations in this region lead to Duchenne muscular dystrophy, a severe genetic muscle wasting disease. Efficient delivery of these gene deletion cargoes is observed in a human cardiomyocyte cell line (AC16), induced pluripotent stem cells, and mesenchymal stem cells.

出版狀態Published - 15 7月 2021


深入研究「Supramolecular Nanosubstrate-Mediated Delivery for CRISPR/Cas9 Gene Disruption and Deletion」主題。共同形成了獨特的指紋。