IHB OpenIR  > 水生生物分子与细胞生物学研究中心  > 期刊论文
Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator
Gu, Qilin1,2; Yang, Xiaojie1,2; He, Xiaozhen1,2; Li, Qing1; Cui, Zongbin1; Cui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China.
2013-10-01
Source PublicationJOURNAL OF GENETICS AND GENOMICS
ISSN1673-8527
Volume40Issue:10Pages:523-531
AbstractConditional 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.; 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.
SubtypeArticle
KeywordZebrafish Transgene Tet-on System Reverse Tet Transactivator Doxycycline
Department[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
DOI10.1016/j.jgg.2013.06.008
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390]
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Genetics & Heredity
WOS SubjectBiochemistry & Molecular Biology ; Genetics & Heredity
WOS IDWOS:000326264300003
WOS KeywordTRANSCRIPTIONAL REGULATION SYSTEMS ; GENE-EXPRESSION ; TET-ON ; CONDITIONAL EXPRESSION ; FUNCTIONAL DISSECTION ; MAMMALIAN-CELLS ; VECTOR SYSTEM ; TIGHT CONTROL ; MICE ; SEQUENCE
Funding OrganizationNational Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390] ; National Basic Research Program of China [2012CB944500]; National Natural Science Foundation of China [31171390]
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Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/19787
Collection水生生物分子与细胞生物学研究中心_期刊论文
Corresponding AuthorCui, ZB (reprint author), Chinese Acad Sci, Key Lab Aquat Biodivers & Conservat, Inst Hydrobiol, Wuhan 430072, Peoples R China.
Affiliation1.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
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,&Cui, ZB .(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.
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