Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells

doi:10.1038/cr.2014.160

	Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells
	Wu, Yuxuan 1,2; Zhou, Hai 1,2,3; Fan, Xiaoying 4; Zhang, Ying 2,5; Zhang, Man 1,2; Wang, Yinghua 1,2; Xie, Zhenfei 1,2; Bai, Meizhu1,2,6 ; Yin, Qi 1,2; Liang, Dan 1,2; Tang, Wei 7; Liao, Jiaoyang 1,2; Zhou, Chikai 1,2; Liu, Wujuan 1,2; Zhu, Ping 4; Guo, Hongshan 4; Pan, Hong 1,2; Wu, Chunlian 3; Shi, Huijuan 8; Wu, Ligang 2,5; Tang, Fuchou 4; Li, Jinsong1,2,6
	2015-01
发表期刊	CELL RESEARCH
ISSN	1001-0602
卷号	25 期号:1 页码:67-79
发表状态	已发表
DOI	10.1038/cr.2014.160
摘要	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.
关键词	CRISPR-Cas9 spermatogonial stem cell gene therapy
收录类别	SCI
语种	英语
资助项目	Shanghai Municipal Commission for Science and Technology[12JC1409600] ; Shanghai Municipal Commission for Science and Technology[13XD1404000] ; Shanghai Municipal Commission for Science and Technology[12JC1409400]
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000349330800009
出版者	INST BIOCHEMISTRY & CELL BIOLOGY
WOS关键词	GERMLINE TRANSMISSION ; IN-VITRO ; MUSCULAR-DYSTROPHY ; CAS9 NUCLEASE ; MODIFIED MICE ; SELF-RENEWAL ; C-KIT ; GENOME ; GENERATION ; CULTURE
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/2336
专题	生命科学与技术学院生命科学与技术学院_特聘教授组_李劲松组生命科学与技术学院_博士生
通讯作者	Li, Jinsong
作者单位	1.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Cell Biol,Grp Epigenet Reprogrammin, Shanghai 200031, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai Key Lab Mol Androl, Shanghai 200031, Peoples R China 3.China West Normal Univ, Coll Life Sci, Nanchong 637002, Sichuan, Peoples R China 4.Peking Univ, Coll Life Sci, Biodynam Opt Imaging Ctr, Key Lab Cell Proliferat & Differentiat,Minist Edu, Beijing 100871, Peoples R China 5.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 6.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai 200031, Peoples R China 7.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Anim Core Facil, Shanghai 200031, Peoples R China 8.Shanghai Inst Planned Parenthood Res, Natl Populat & Family Planning Comm, Key Lab Contracept Drugs & Devices, Shanghai 200032, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Wu, Yuxuan,Zhou, Hai,Fan, Xiaoying,et al. Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells[J]. CELL RESEARCH,2015,25(1):67-79.
APA	Wu, Yuxuan.,Zhou, Hai.,Fan, Xiaoying.,Zhang, Ying.,Zhang, Man.,...&Li, Jinsong.(2015).Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells.CELL RESEARCH,25(1),67-79.
MLA	Wu, Yuxuan,et al."Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells".CELL RESEARCH 25.1(2015):67-79.