| Title | Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells |
| Authors | Wu, Yuxuan Zhou, Hai Fan, Xiaoying Zhang, Ying Zhang, Man Wang, Yinghua Xie, Zhenfei Bai, Meizhu Yin, Qi Liang, Dan Tang, Wei Liao, Jiaoyang Zhou, Chikai Liu, Wujuan Zhu, Ping Guo, Hongshan Pan, Hong Wu, Chunlian Shi, Huijuan Wu, Ligang Tang, Fuchou Li, Jinsong |
| Affiliation | Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Cell Biol,Grp Epigenet Reprogrammin, Shanghai 200031, Peoples R China. Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai Key Lab Mol Androl, Shanghai 200031, Peoples R China. China West Normal Univ, Coll Life Sci, Nanchong 637002, Sichuan, Peoples R China. Peking Univ, Coll Life Sci, Biodynam Opt Imaging Ctr, Key Lab Cell Proliferat & Differentiat,Minist Edu, Beijing 100871, Peoples R China. Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai Key Lab Mol Androl,State Key Lab Mol Bio, Shanghai 200031, Peoples R China. Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai 200031, Peoples R China. Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Anim Core Facil, Shanghai 200031, Peoples R China. Shanghai Inst Planned Parenthood Res, Natl Populat & Family Planning Comm, Key Lab Contracept Drugs & Devices, Shanghai 200032, Peoples R China. |
| Keywords | CRISPR-Cas9 spermatogonial stem cell gene therapy GERMLINE TRANSMISSION IN-VITRO MUSCULAR-DYSTROPHY CAS9 NUCLEASE MODIFIED MICE SELF-RENEWAL C-KIT GENOME GENERATION CULTURE |
| Issue Date | 2015 |
| Publisher | 细胞研究英文版 |
| Citation | CELL RESEARCH.2015,25,(1),67-79. |
| Abstract | Spermatogonial stem cells (SSCs) can produce numerous male gametes after transplantation into recipient testes, presenting a valuable approach for gene therapy and continuous production of gene-modified animals. However, successful genetic manipulation of SSCs has been limited, partially due to complexity and low efficiency of currently available genetic editing techniques. Here, we show that efficient genetic modifications can be introduced into SSCs using the CRISPR-Cas9 system. We used the CRISPR-Cas9 system to mutate an EGFP transgene or the endogenous Crygc gene in SCCs. The mutated SSCs underwent spermatogenesis after transplantation into the seminiferous tubules of infertile mouse testes. Round spermatids were generated and, after injection into mature oocytes, supported the production of heterozygous offspring displaying the corresponding mutant phenotypes. Furthermore, a disease-causing mutation in Crygc (Crygc(-/-)) that pre-existed in SSCs could be readily repaired by CRISPR-Cas9-induced nonhomologous end joining (NHEJ) or homology-directed repair (HDR), resulting in SSC lines carrying the corrected gene with no evidence of off-target modifications as shown by whole-genome sequencing. Fertilization using round spermatids generated from these lines gave rise to offspring with the corrected phenotype at an efficiency of 100%. Our results demonstrate efficient gene editing in mouse SSCs by the CRISPR-Cas9 system, and provide the proof of principle of curing a genetic disease via gene correction in SSCs. |
| URI | http://hdl.handle.net/20.500.11897/341768 |
| ISSN | 1001-0602 |
| DOI | 10.1038/cr.2014.160 |
| Indexed | SCI(E) PubMed 中国科技核心期刊(ISTIC) 中国科学引文数据库(CSCD) |
| Appears in Collections: | 生命科学学院 细胞增殖分化调控机理研究教育部重点实验室 |
