Knowledge Management System of Northwest Institute of Plateau Biology, CAS
| A suicidal zinc finger nuclease expression coupled with a surrogate reporter for efficient genome engineering | |
| Zhang, Cunfang1,2; Xu, Kun1; Hu, Linyong2; Wang, Ling1,3; Zhang, Tingting1; Ren, Chonghua1; Zhang, Zhiying1; Zhang, ZY (reprint author), Northwest A&F Univ, Coll Anim Sci & Technol, 22 Xinong Rd, Yangling 712100, Shaanxi, Peoples R China. | |
| 2015-02-01 | |
| 发表期刊 | BIOTECHNOLOGY LETTERS
 |
| 卷号 | 37期号:2页码:299-305 |
| 文章类型 | Article |
| 摘要 | Genome editing with engineered nucleases, such as zinc-finger nucleases (ZFNs) and TALE nucleases, remains confronted with a high risk of cellular toxicity induced by off-targeting. Here we describe the construction of a suicidal nuclease expression vector in which a pair of ZFNs genes were flanked of its target sites. To further enrich the targeted cells, the suicidal ZFN expression cassette was also inserted within an eGFP reporter, to disrupt the ORF of eGFP gene. ZFN-associated toxicity was reduced by similar to 40 % with this new system, and the activities of ZFNs were similar to 4.5 % lower. We conclude that using this new suicidal ZFN expression and surrogate reporter system represents an improvement for genomic editing by reducing toxicity and allowing easy detection of edited cells by eGFP analysis.; Genome editing with engineered nucleases, such as zinc-finger nucleases (ZFNs) and TALE nucleases, remains confronted with a high risk of cellular toxicity induced by off-targeting. Here we describe the construction of a suicidal nuclease expression vector in which a pair of ZFNs genes were flanked of its target sites. To further enrich the targeted cells, the suicidal ZFN expression cassette was also inserted within an eGFP reporter, to disrupt the ORF of eGFP gene. ZFN-associated toxicity was reduced by similar to 40 % with this new system, and the activities of ZFNs were similar to 4.5 % lower. We conclude that using this new suicidal ZFN expression and surrogate reporter system represents an improvement for genomic editing by reducing toxicity and allowing easy detection of edited cells by eGFP analysis. |
| 关键词 | Cytotoxicity Double-strand-breaks Gene Targeting Homologous Recombination Off-targeting Zinc Finger Nuclease |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 关键词[WOS] | HOMOLOGOUS RECOMBINATION ; MAMMALIAN-CELLS ; GENE ; DROSOPHILA |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Biotechnology & Applied Microbiology |
| WOS类目 | Biotechnology & Applied Microbiology |
| WOS记录号 | WOS:000349230800006 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://210.75.249.4/handle/363003/5457 |
| 专题 | 中国科学院西北高原生物研究所 |
| 通讯作者 | Zhang, ZY (reprint author), Northwest A&F Univ, Coll Anim Sci & Technol, 22 Xinong Rd, Yangling 712100, Shaanxi, Peoples R China. |
| 作者单位 | 1.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Shaanxi, Peoples R China 2.Chinese Acad Sci, Key Lab Adaptat & Evolut Plateau Biota, Northwest Inst Plateau Biol, Xining, Peoples R China 3.Shaanxi Univ Technol, Sch Biol Sci Technol & Engn, Hanzhong, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Cunfang,Xu, Kun,Hu, Linyong,et al. A suicidal zinc finger nuclease expression coupled with a surrogate reporter for efficient genome engineering[J]. BIOTECHNOLOGY LETTERS,2015,37(2):299-305. |
| APA | Zhang, Cunfang.,Xu, Kun.,Hu, Linyong.,Wang, Ling.,Zhang, Tingting.,...&Zhang, ZY .(2015).A suicidal zinc finger nuclease expression coupled with a surrogate reporter for efficient genome engineering.BIOTECHNOLOGY LETTERS,37(2),299-305. |
| MLA | Zhang, Cunfang,et al."A suicidal zinc finger nuclease expression coupled with a surrogate reporter for efficient genome engineering".BIOTECHNOLOGY LETTERS 37.2(2015):299-305. |
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