Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish
Liu, Chunyan1,2,3; Song, Guili1,2; Mao, Lin1,2,3; Long, Yong1,2; Li, Qing1,2; Cui, Zongbin1,2
刊名PLOS ONE
2015-10-05
卷号10期号:10
ISSN号1932-6203
英文摘要Enhancer trapping (ET) is a powerful approach to establish tissue-or cell-specific reporters and identify expression patterns of uncharacterized genes. Although a number of enhancer-trapping vectors have been developed and a large library of fish lines with distinct tissue- or cell-specific expression of reporter genes have been generated, the specificity and efficiency of trapping vectors need to be improved because of the bias interaction of minimal promoters with genomic enhancers. Accordingly, we generated an enhancer-trapping vector pTME that contains a minimal mouse metallothionein gene (mMTI) promoter upstream of EGFP reporter. In the first round of screening, twelve zebrafish lines that carry a single copy of ET cassettes were characterized to have tissue-or cell-specific EGFP expression. One of the highly conserved noncoding elements near an insertion site of trapping cassettes was characterized as an enhancer that can specifically regulate the expression of EGFP in cells of the central nervous system. In addition, the pTME vector contains a mutation-cassette that is able to effectively block the transcription of an endogenous gene in an ET line with ubiquitous EGFP expression. Thus, the pTME vector can be used as an alternative tool for both enhancer trapping and mutagenesis across a target genome.
WOS标题词Science & Technology
类目[WOS]Multidisciplinary Sciences
研究领域[WOS]Science & Technology - Other Topics
关键词[WOS]CONSERVED NONCODING SEQUENCES ; SLEEPING-BEAUTY TRANSPOSON ; IN-SITU HYBRIDIZATION ; TRANSCRIPTION FACTOR ; GENE-EXPRESSION ; GENOME ; VIVO ; DROSOPHILA ; DISSECTION ; MUTATIONS
收录类别SCI
语种英语
WOS记录号WOS:000362499200036
内容类型期刊论文
源URL[http://ir.ihb.ac.cn/handle/342005/27272]  
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, Wuhan 430072, Hubei, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
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Liu, Chunyan,Song, Guili,Mao, Lin,et al. Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish[J]. PLOS ONE,2015,10(10).
APA Liu, Chunyan,Song, Guili,Mao, Lin,Long, Yong,Li, Qing,&Cui, Zongbin.(2015).Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish.PLOS ONE,10(10).
MLA Liu, Chunyan,et al."Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish".PLOS ONE 10.10(2015).
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