Sleeping beauty transposon-mediated poly(A)-trapping and insertion mutagenesis in mouse embryonic stem cells
文献类型：期刊论文
| 作者 | Cui, Zongbin3; Zhong, Shan1,2,4; Li, Qing3; Ren, Jing3; Song, Guili3; Zhao, Yi4 |
| 刊名 | ENVIRONMENTAL AND MOLECULAR MUTAGENESIS
 |
| 出版日期 | 2018-10-01 |
| 卷号 | 59期号:8页码:687-697 |
| 关键词 | mouse embryonic stem cells sleeping beauty transposon poly(A) trapping insertion mutagenesis |
| ISSN号 | 0893-6692 |
| DOI | 10.1002/em.22234 |
| 英文摘要 | Saturation mutagenesis of all endogenous genes within the mouse genome remains a challenging task, although a plenty of gene-editing approaches are available for this purpose. Here, a poly(A)-trap vector was generated for insertion mutagenesis in mouse embryonic stem (mES) cells. This vector contains an expression cassette of neomycin (Neo)-resistant gene lacking a poly(A) signal and flanked by two inverted terminal repeats of the Sleeping Beauty (SB) transposon. The whole poly(A)-trap cassette can transpose into target TA dinucleotides, properly splice with endogenous genes and effectively interrupt the transcription of trapped genes in mES cells after transient induction of SB expression by doxycycline (DOX)-treatment at 1 g/ml, leading to the formation of multiple geneticin (G418)-resistant cell clones. In the first round of mutation screening, we identified six transposition events from 23 cell clones, including four inserted into an endogenous gene and two landed between endogenous genes. The abilities of self-renewal, totipotency, genetic stability and differentiation of syngap1(+/-) cells were not affected by DOX-treatment and G418-selection. These findings suggest that this SB transposon-mediated poly(A)-trap vector can be used as an alternative tool for a large-scale screening of mES cells with a gene mutation and for further generation of mutant mouse strains. Environ. Mol. Mutagen. 59:687-697, 2018. (c) 2018 Wiley Periodicals, Inc. |
| WOS研究方向 | Environmental Sciences & Ecology ; Genetics & Heredity ; Toxicology |
| 语种 | 英语 |
| WOS记录号 | WOS:000447275100002 |
| 出版者 | WILEY |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/6006]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Cui, Zongbin; Zhong, Shan |
| 作者单位 | 1.Hubei Prov Key Lab Allergy & Immunol, Wuhan, Hubei, Peoples R China 2.Hubei Prov Key Lab Dev Originated Dis, Wuhan, Hubei, Peoples R China 3.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Hubei, Peoples R China 4.Wuhan Univ, Sch Basic Med Sci, Dept Genet, Wuhan, Hubei, Peoples R China |
| 推荐引用方式 GB/T 7714 | Cui, Zongbin,Zhong, Shan,Li, Qing,et al. Sleeping beauty transposon-mediated poly(A)-trapping and insertion mutagenesis in mouse embryonic stem cells[J]. ENVIRONMENTAL AND MOLECULAR MUTAGENESIS,2018,59(8):687-697. |
| APA | Cui, Zongbin,Zhong, Shan,Li, Qing,Ren, Jing,Song, Guili,&Zhao, Yi.(2018).Sleeping beauty transposon-mediated poly(A)-trapping and insertion mutagenesis in mouse embryonic stem cells.ENVIRONMENTAL AND MOLECULAR MUTAGENESIS,59(8),687-697. |
| MLA | Cui, Zongbin,et al."Sleeping beauty transposon-mediated poly(A)-trapping and insertion mutagenesis in mouse embryonic stem cells".ENVIRONMENTAL AND MOLECULAR MUTAGENESIS 59.8(2018):687-697. |
入库方式： OAI收割
来源：武汉植物园
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