Efficient bi-allelic gene knockout and site-specific knock-in mediated   by TALENs in pigs

doi:10.1038/srep06926

CORC > 北京大学 > 工学院

	Efficient bi-allelic gene knockout and site-specific knock-in mediated by TALENs in pigs
	Yao, Jing ; Huang, Jiaojiao ; Hai, Tang ; Wang, Xianlong ; Qin, Guosong ; Zhang, Hongyong ; Wu, Rong ; Cao, Chunwei ; Xi, Jianzhong Jeff ; Yuan, Zengqiang ; Zhao, Jianguo
刊名	scientific reports
	2014
关键词	ZINC-FINGER NUCLEASES FIBROBLAST-GROWTH-FACTOR PARKIN-DEFICIENT MICE EMBRYO MICROINJECTION NULL MICE EFFECTOR NUCLEASES TARGETING DNA GENERATION MUTATIONS DISEASE
DOI	10.1038/srep06926
英文摘要	Pigs are ideal organ donors for xenotransplantation and an excellent model for studying human diseases, such as neurodegenerative disease. Transcription activator-like effector nucleases (TALENs) are used widely for gene targeting in various model animals. Here, we developed a strategy using TALENs to target the GGTA1, Parkin and DJ-1 genes in the porcine genome using Large White porcine fibroblast cells without any foreign gene integration. In total, 5% (2/40), 2.5% (2/80), and 22% (11/50) of the obtained colonies of fibroblast cells were mutated for GGTA1, Parkin, and DJ-1, respectively. Among these mutant colonies, over 1/3 were bi-allelic knockouts (KO), and no off-target cleavage was detected. We also successfully used single-strand oligodeoxynucleotides to introduce a short sequence into the DJ-1 locus. Mixed DJ-1 mutant colonies were used as donor cells for somatic cell nuclear transfer (SCNT), and three female piglets were obtained (two were bi-allelically mutated, and one was mono-allelically mutated). Western blot analysis showed that the expression of the DJ-1 protein was disrupted in KO piglets. These results imply that a combination of TALENs technology with SCNT can efficiently generate bi-allelic KO pigs without the integration of exogenous DNA. These DJ-1 KO pigs will provide valuable information for studying Parkinson's disease.; Multidisciplinary Sciences; SCI(E); PubMed; 3; ARTICLE; Zhaojg@ioz.ac.cn; 6926; 4
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/188564]
专题	工学院
推荐引用方式 GB/T 7714	Yao, Jing,Huang, Jiaojiao,Hai, Tang,et al. Efficient bi-allelic gene knockout and site-specific knock-in mediated by TALENs in pigs[J]. scientific reports,2014.
APA	Yao, Jing.,Huang, Jiaojiao.,Hai, Tang.,Wang, Xianlong.,Qin, Guosong.,...&Zhao, Jianguo.(2014).Efficient bi-allelic gene knockout and site-specific knock-in mediated by TALENs in pigs.scientific reports.
MLA	Yao, Jing,et al."Efficient bi-allelic gene knockout and site-specific knock-in mediated by TALENs in pigs".scientific reports (2014).