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题名: Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator
作者: Gu, Qilin1, 2; Yang, Xiaojie1, 2; He, Xiaozhen1, 2; Li, Qing1; Cui, Zongbin1
通讯作者: Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China.
关键词: Zebrafish ; Transgene ; Tet-on system ; Reverse Tet transactivator ; Doxycycline
刊名: JOURNAL OF GENETICS AND GENOMICS
发表日期: 2013-10-01
DOI: 10.1016/j.jgg.2013.06.008
卷: 40, 期:10, 页:523-531
收录类别: SCI
文章类型: Article
部门归属: [Gu, Qilin ; Yang, Xiaojie ; He, Xiaozhen ; Li, Qing ; Cui, Zongbin] Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China ; [Gu, Qilin ; Yang, Xiaojie ; He, Xiaozhen] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390]
类目[WOS]: Biochemistry & Molecular Biology ; Genetics & Heredity
研究领域[WOS]: Biochemistry & Molecular Biology ; Genetics & Heredity
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.
关键词[WOS]: TRANSCRIPTIONAL REGULATION SYSTEMS ; GENE-EXPRESSION ; TET-ON ; CONDITIONAL EXPRESSION ; FUNCTIONAL DISSECTION ; MAMMALIAN-CELLS ; VECTOR SYSTEM ; TIGHT CONTROL ; MICE ; SEQUENCE
语种: 英语
WOS记录号: WOS:000326264300003
ISSN号: 1673-8527
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/19787
Appears in Collections:水生生物分子与细胞生物学研究中心_期刊论文

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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

Recommended Citation:
Gu, Qilin; Yang, Xiaojie; He, Xiaozhen; Li, Qing; Cui, Zongbin.Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator,JOURNAL OF GENETICS AND GENOMICS,2013,40(10):523-531
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