文献类型： 外文期刊

作者： Ban, Qian ¹ ; Yang, Peng ² ; Chou, Shih-Jie ³ ; Qiao, Li ¹ ; Xia, Haidong ¹ ; Xue, Jingjing ² ; Wang, Fang ⁵ ; Xu, Xiaobi ¹ ;

作者机构： 1.Anhui Univ, Ctr Stem Cell & Translat Med, Sch Life Sci, Hefei 230601, Peoples R China

2.Univ Calif Los Angeles, David Geffen Sch Med, Crump Inst Mol Imaging CIMI, Calif NanoSyst Inst CNSI,Dept Mol & Med Pharmacol, Los Angeles, CA 90095 USA

3.Natl Yang Ming Chiao Tung Univ, Taipei Vet Gen Hosp, Coll Med, Inst Pharmacol,Dept Med Res, 155,Sec 2,Linong St 112, Taipei, Taiwan

4.Natl Yang Ming Chiao Tung Univ, Taipei Vet Gen Hosp, Coll Med, Inst Pharmacol,Stem Cell Ctr,Div Basic Res, 155,Sec 2,Linong St 112, Taipei, Taiwan

5.Fudan Univ, Dept Macromol Sci, State Key Lab Mol Engn Polymers, Shanghai 200433, Peoples R China

6.Univ Calif Los Angeles, Calif NanoSyst Inst CNSI, Dept Chem & Biochem, Dept Bioengn,Dept Mat Sci & Engn, Los Angeles, CA 90095 USA

7.Tongji Univ, Sch Mat Sci & Engn, 1239 Siping Rd, Shanghai 200092, Peoples R China

8.Natl Chung Hsing Univ NCHU, I Ctr Adv Sci & Technol iCAST, Innovat & Dev Ctr Sustainable Agr, Dept Chem, 145 Xingda Rd, Taichung 402, Taiwan

9.Anhui Acad Agr Sci, Inst Anim Husb & Vet Med, Anhui Prov Key Lab Livestock & Poultry Prod Safet, Hefei 230031, Peoples R China

10.Foshan Univ, Sch Stomatol & Med, Foshan 528000, Peoples R China

11.Univ Calif Los Angeles, Eli & Edythe Broad Ctr Regenerat Med & Stem Cell, David Geffen Sch Med,Childrens Discovery & Innova, Calif NanoSyst Inst CNSI,Dept Pediat, Los Angeles, CA 90095 USA

关键词： CRISPR; Cas9; Duchenne muscular dystrophy; gene editing; nanosubstrate-mediated delivery; supramolecular nanoparticles

期刊名称：SMALL （ 影响因子：13.281； 五年影响因子：12.463 ）

ISSN： 1613-6810

年卷期： 2021 年 17 卷 28 期

页码：

收录情况： SCI

摘要： 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 center dot sgRNA ribonucleoprotein (RNP) complexes can be encapsulated into supramolecular nanoparticles (SMNP) to form RNP subset of 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.