Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator
文献类型：期刊论文
| 作者 | Gu, Qilin1,2; Yang, Xiaojie1,2; He, Xiaozhen1,2; Li, Qing1; Cui, Zongbin1 |
| 刊名 | JOURNAL OF GENETICS AND GENOMICS
 |
| 出版日期 | 2013-10-01 |
| 卷号 | 40期号:10页码:523-531 |
| 关键词 | Zebrafish Transgene Tet-on system Reverse Tet transactivator Doxycycline |
| ISSN号 | 1673-8527 |
| 通讯作者 | Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China. |
| 中文摘要 | Conditional expression of a target gene during zebrafish development is a powerful approach to elucidate gene functions. The tetracycline-controlled systems have been successfully used in the modulation of gene expression in mammalian cells, but few lines of zebrafish carrying these systems are currently available. In this study, we had generated a stable transgenic zebrafish line that ubiquitously expressed the second-generation of reverse Tet transactivator (rtTA-M2). Southern blotting analysis and high-throughput genome sequencing verified that a single copy of rtTA-M2 gene had stably integrated into the zebrafish genome. After induction with doxycycline (Dox), a strong green fluorescent protein (GFP) was seen in rtTA-transgenic eggs injected with pTRE-EGFP plasmids. The fluorescent signal gradually decreased after the withdrawal of Dox and disappeared. However, leaky expression of GFP was undetectable before Dox-induction. Additionally, transgenic embryos expressing rtTA-M2 exhibited no obvious defects in morphological phenotypes, hatching behavior and expression patterns of developmental marker genes, suggesting that rtTA-M2 had little effect on the development of transgenic zebrafish. Moreover, expressed Dickkopf-1 (DKK1) in pTRE-DKK1-injected embryos led to alterations in the expression of marker genes associated with Wnt signaling. Thus, this rtTA-transgenic zebrafish can be utilized to dissect functions of genes in a temporal manner. |
| 英文摘要 | Conditional expression of a target gene during zebrafish development is a powerful approach to elucidate gene functions. The tetracycline-controlled systems have been successfully used in the modulation of gene expression in mammalian cells, but few lines of zebrafish carrying these systems are currently available. In this study, we had generated a stable transgenic zebrafish line that ubiquitously expressed the second-generation of reverse Tet transactivator (rtTA-M2). Southern blotting analysis and high-throughput genome sequencing verified that a single copy of rtTA-M2 gene had stably integrated into the zebrafish genome. After induction with doxycycline (Dox), a strong green fluorescent protein (GFP) was seen in rtTA-transgenic eggs injected with pTRE-EGFP plasmids. The fluorescent signal gradually decreased after the withdrawal of Dox and disappeared. However, leaky expression of GFP was undetectable before Dox-induction. Additionally, transgenic embryos expressing rtTA-M2 exhibited no obvious defects in morphological phenotypes, hatching behavior and expression patterns of developmental marker genes, suggesting that rtTA-M2 had little effect on the development of transgenic zebrafish. Moreover, expressed Dickkopf-1 (DKK1) in pTRE-DKK1-injected embryos led to alterations in the expression of marker genes associated with Wnt signaling. Thus, this rtTA-transgenic zebrafish can be utilized to dissect functions of genes in a temporal manner. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemistry & Molecular Biology ; Genetics & Heredity |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Genetics & Heredity |
| 关键词[WOS] | TRANSCRIPTIONAL REGULATION SYSTEMS ; GENE-EXPRESSION ; TET-ON ; CONDITIONAL EXPRESSION ; FUNCTIONAL DISSECTION ; MAMMALIAN-CELLS ; VECTOR SYSTEM ; TIGHT CONTROL ; MICE ; SEQUENCE |
| 收录类别 | SCI |
| 资助信息 | National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] |
| 语种 | 英语 |
| WOS记录号 | WOS:000326264300003 |
| 公开日期 | 2014-01-24 |
| 源URL | [http://ir.ihb.ac.cn/handle/342005/19787]  |
| 专题 | 水生生物研究所_水生生物分子与细胞生物学研究中心_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Gu, Qilin,Yang, Xiaojie,He, Xiaozhen,et al. Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator[J]. JOURNAL OF GENETICS AND GENOMICS,2013,40(10):523-531. |
| APA | Gu, Qilin,Yang, Xiaojie,He, Xiaozhen,Li, Qing,&Cui, Zongbin.(2013).Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator.JOURNAL OF GENETICS AND GENOMICS,40(10),523-531. |
| MLA | Gu, Qilin,et al."Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator".JOURNAL OF GENETICS AND GENOMICS 40.10(2013):523-531. |
入库方式： OAI收割
来源：水生生物研究所
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