Rationally designed nanoparticle delivery of Cas9 ribonucleoprotein for effective gene editing

被引:14
作者
Chae, Se-Youl [1 ]
Jeong, Euihwan [1 ,3 ]
Kang, Seounghun [1 ]
Yim, Yeajee [1 ]
Kim, Jin-Soo [3 ]
Min, Dal-Hee [1 ,2 ]
机构
[1] Seoul Natl Univ, Dept Chem, Seoul 08826, South Korea
[2] Lemonex Inc, Inst BioTherapeut Convergence Technol, Seoul 06683, South Korea
[3] Inst Basic Sci IBS, Ctr Genome Engn, Seoul, South Korea
基金
新加坡国家研究基金会;
关键词
Cas9; ribonucleoprotein; Gene knockout; Genome editing; Myostatin; Porous nanoparticle; MESOPOROUS SILICA NANOPARTICLES; MOUSE MODEL; EFFICIENT DELIVERY; HUMAN-CELLS; RNA; DNA; PROTEIN; ENDONUCLEASE; CRISPR; SIZES;
D O I
10.1016/j.jconrel.2022.02.035
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Programmable endonucleases such as CRISPR/Cas9 system emerge as a promising tool to treat genetic and non genetic diseases such as hypercholesterolemia, Duchenne muscular dystrophy, and cancer. However, the lack of safe and efficient vehicles that enable intracellular delivery of CRISPR/Cas9 endonuclease is a big hurdle for its therapeutic applications. Here, we employed porous nanoparticle for the Cas9 ribonucleoprotein (RNP) delivery and achieved efficient knockout of target genes in vitro and in vivo. The porous nanoparticle, called 'BALL', enabled safe and direct intracellular Cas9 RNP delivery by improving bioavailability and serum stability. The BALL-mediated delivery of Cas9 RNP showed superior indel efficiency of about 40% in vitro and 20% in vivo in a model system employing green fluorescent protein (GFP). More importantly, intramuscular injection of the Cas9 RNP-BALL complex targeting the myostatin (MSTN) gene which is known to suppress muscle growth achieved successful knockout of the MSTN gene, resulting in the increase of muscle and the improved motor functions. Thus, we believe that the BALL is a promising delivery system for CRISPR-based genome editing technology, which can be applied to the treatment of various genetic diseases.
引用
收藏
页码:108 / 119
页数:12
相关论文
共 46 条
[1]   Myostatin gene promoter: structure, conservation and importance as a target for muscle modulation [J].
Carvalho Grade, Carla Vermeulen ;
Mantovani, Carolina Stefano ;
Alvares, Lucia Elvira .
JOURNAL OF ANIMAL SCIENCE AND BIOTECHNOLOGY, 2019, 10 (1)
[2]   Extra-Large Pore Mesoporous Silica Nanoparticles Enabling Co-Delivery of High Amounts of Protein Antigen and Toll-like Receptor 9 Agonist for Enhanced Cancer Vaccine Efficacy [J].
Cha, Bong Geun ;
Jeong, Ji Hoon ;
Kim, Jaeyun .
ACS CENTRAL SCIENCE, 2018, 4 (04) :484-492
[3]   Heritable Gene Knockout in Caenorhabditis elegans by Direct Injection of Cas9-sgRNA Ribonucleoproteins [J].
Cho, Seung Woo ;
Lee, Jihyun ;
Carroll, Dana ;
Kim, Jin-Soo ;
Lee, Junho .
GENETICS, 2013, 195 (03) :1177-+
[4]   Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease [J].
Cho, Seung Woo ;
Kim, Sojung ;
Kim, Jong Min ;
Kim, Jin-Soo .
NATURE BIOTECHNOLOGY, 2013, 31 (03) :230-232
[5]   Surface pH buffering to promote degradation of mesoporous silica nanoparticles under a physiological condition [J].
Choi, Eunshil ;
Kim, Sehoon .
JOURNAL OF COLLOID AND INTERFACE SCIENCE, 2019, 533 :463-470
[6]   Multiplex Genome Engineering Using CRISPR/Cas Systems [J].
Cong, Le ;
Ran, F. Ann ;
Cox, David ;
Lin, Shuailiang ;
Barretto, Robert ;
Habib, Naomi ;
Hsu, Patrick D. ;
Wu, Xuebing ;
Jiang, Wenyan ;
Marraffini, Luciano A. ;
Zhang, Feng .
SCIENCE, 2013, 339 (6121) :819-823
[7]   Therapeutic genome editing: prospects and challenges [J].
Cox, David Benjamin Turitz ;
Platt, Randall Jeffrey ;
Zhang, Feng .
NATURE MEDICINE, 2015, 21 (02) :121-131
[8]   CRISPR-Cas9 In Vivo Gene Editing for Transthyretin Amyloidosis [J].
Gillmore, Julian D. ;
Gane, Ed ;
Taubel, Jorg ;
Kao, Justin ;
Fontana, Marianna ;
Maitland, Michael L. ;
Seitzer, Jessica ;
O'Connell, Daniel ;
Walsh, Kathryn R. ;
Wood, Kristy ;
Phillips, Jonathan ;
Xu, Yuanxin ;
Amaral, Adam ;
Boyd, Adam P. ;
Cehelsky, Jeffrey E. ;
McKee, Mark D. ;
Schiermeier, Andrew ;
Harari, Olivier ;
Murphy, Andrew ;
Kyratsous, Christos A. ;
Zambrowicz, Brian ;
Soltys, Randy ;
Gutstein, David E. ;
Leonard, John ;
Sepp-Lorenzino, Laura ;
Lebwohl, David .
NEW ENGLAND JOURNAL OF MEDICINE, 2021, 385 (06) :493-502
[9]   Tailoring nanocarriers for intracellular protein delivery [J].
Gu, Zhen ;
Biswas, Anuradha ;
Zhao, Muxun ;
Tang, Yi .
CHEMICAL SOCIETY REVIEWS, 2011, 40 (07) :3638-3655
[10]   Direct cellular delivery of human proteasomes to delay tau aggregation [J].
Han, Dong Hoon ;
Na, Hee-Kyung ;
Choi, Won Hoon ;
Lee, Jung Hoon ;
Kim, Yun Kyung ;
Won, Cheolhee ;
Lee, Seung-Han ;
Kim, Kwang Pyo ;
Kuret, Jeff ;
Min, Dal-Hee ;
Lee, Min Jae .
NATURE COMMUNICATIONS, 2014, 5