Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer

	Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer
	Bao, L; Chen, HD; Jong, UM; Rim, CH; Li, WL; Lin, XJ; Zhang, D; Luo, Q; Cui, C; Huang, HF
刊名	SCIENCE CHINA-LIFE SCIENCES
	2014
卷号	57 期号:2 页码:263-268
关键词	pig xenotransplantation ZFNs GGTA1 biallelic knockout SCNT
通讯作者	Fu, ZX (reprint author), Zhejiang Univ, Sch Med, Coll Anim Sci, Hangzhou 310058, Zhejiang, Peoples R China.,fzxljl@163.com
英文摘要	Genetically modified pigs are valuable models of human disease and donors of xenotransplanted organs. Conventional gene targeting in pig somatic cells is extremely inefficient. Zinc-finger nuclease (ZFN) technology has been shown to be a powerful tool for efficiently inducing mutations in the genome. However, ZFN-mediated targeting in pigs has rarely been achieved. Here, we used ZFNs to knock out the porcine alpha-1, 3-galactosyl-transferase (GGTA1) gene, which generates Gal epitopes that trigger hyperacute immune rejection in pig-to-human transplantation. Primary pig fibroblasts were transfected with ZFNs targeting the coding region of GGTA1. Eighteen mono-allelic and four biallelic knockout cell clones were obtained after drug selection with efficiencies of 23.4% and 5.2%, respectively. The biallelic cells were used to produce cloned pigs via somatic cell nuclear transfer (SCNT). Three GGTA1 null piglets were born, and one knockout primary fibroblast cell line was established from a cloned fetus. Gal epitopes on GGTA1 null pig cells were completely eliminated from the cell membrane. Functionally, GGTA1 knockout cells were protected from complement-mediated immune attacks when incubated with human serum. This study demonstrated that ZFN is an efficient tool in creating gene-modified pigs. GGTA1 null pigs and GGTA1 null fetal fibroblasts would benefit research and pig-to-human transplantation.
学科主题	Life Sciences & Biomedicine - Other Topics
类目[WOS]	Biology
关键词[WOS]	STEM-CELLS ; GENE ; XENOTRANSPLANTATION ; BABOONS ; ANTIBODIES ; CLONING ; SYSTEM ; DONORS ; MODEL
收录类别	SCI
语种	英语
WOS记录号	WOS:000331206000015
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/147]
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	Bao, L,Chen, HD,Jong, UM,et al. Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer[J]. SCIENCE CHINA-LIFE SCIENCES,2014,57(2):263-268.
APA	Bao, L.,Chen, HD.,Jong, UM.,Rim, CH.,Li, WL.,...&Fu, ZX.(2014).Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer.SCIENCE CHINA-LIFE SCIENCES,57(2),263-268.
MLA	Bao, L,et al."Generation of GGTA1 biallelic knockout pigs via zinc-finger nucleases and somatic cell nuclear transfer".SCIENCE CHINA-LIFE SCIENCES 57.2(2014):263-268.