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

	Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells
	Wu, YX; Zhou, H; Fan, XY; Zhang, Y; Zhang, M; Wang, YH; Xie, ZF; Bai, MZ; Yin, Q; Liang, D
刊名	CELL RESEARCH
	2015
卷号	25 期号:1 页码:67-79
关键词	CRISPR-Cas9 spermatogonial stem cell gene therapy
通讯作者	Li, JS (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Cell Biol,Grp Epigenet Reprogrammin, Shanghai 200031, Peoples R China.,lgwu@sibcb.ac.cn ; tangfuchou@pku.edu.cn ; jsli@sibcb.ac.cn
英文摘要	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.
学科主题	Cell Biology
类目[WOS]	Cell Biology
关键词[WOS]	GERMLINE TRANSMISSION ; IN-VITRO ; MUSCULAR-DYSTROPHY ; CAS9 NUCLEASE ; MODIFIED MICE ; SELF-RENEWAL ; C-KIT ; GENOME ; GENERATION ; CULTURE
收录类别	SCI
语种	英语
WOS记录号	WOS:000349330800009
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/9]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Wu, YX,Zhou, H,Fan, XY,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, YX.,Zhou, H.,Fan, XY.,Zhang, Y.,Zhang, M.,...&Li, JS.(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, YX,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.