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    Enhancer trapping in zebrafish using the Sleeping Beauty transposon

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    Name:
    Enhancer trapping in zebrafish ...
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    Genre
    Review
    Journal
    Date
    2004-09-03
    Author
    Balciunas, D
    Davidson, AE
    Sivasubbu, S
    Hermanson, SB
    Welle, Z
    Ekker, SC
    Subject
    Animals
    DNA Transposable Elements
    Embryonic Development
    Enhancer Elements, Genetic
    Gene Transfer Techniques
    Genomics
    Germ Cells
    Glycoside Hydrolases
    Green Fluorescent Proteins
    In Situ Hybridization
    Motor Neurons
    Pilot Projects
    Polymerase Chain Reaction
    Promoter Regions, Genetic
    Sequence Analysis, DNA
    Zebrafish
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    Permanent link to this record
    http://hdl.handle.net/20.500.12613/5656
    
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    DOI
    10.1186/1471-2164-5-62
    Abstract
    Background: Among functional elements of a metazoan gene, enhancers are particularly difficult to find and annotate. Pioneering experiments in Drosophila have demonstrated the value of enhancer "trapping" using an invertebrate to address this functional genomics problem. Results: We modulated a Sleeping Beauty transposon-based transgenesis cassette to establish an enhancer trapping technique for use in a vertebrate model system, zebrafish Danio rerio. We established 9 lines of zebrafish with distinct tissue- or organ-specific GFP expression patterns from 90 founders that produced GFP-expressing progeny. We have molecularly characterized these lines and show that in each line, a specific GFP expression pattern is due to a single transposition event. Many of the insertions are into introns of zebrafish genes predicted in the current genome assembly. We have identified both previously characterized as well as novel expression patterns from this screen. For example, the ET7 line harbors a transposon insertion near the mkp3 locus and expresses GFP in the midbrain-hindbrain boundary, forebrain and the ventricle, matching a subset of the known FGF8-dependent mkp3 expression domain. The ET2 line, in contrast, expresses GFP specifically in caudal primary motoneurons due to an insertion into the poly(ADPribose) glycohydrolase (PARG) locus. This surprising expression pattern was confirmed using in situ hybridization techniques for the endogenous PARG mRNA, indicating the enhancer trap has replicated this unexpected and highly localized PARG expression with good fidelity. Finally, we show that it is possible to excise a Sleeping Beauty transposon from a genomic location in the zebrafish germline. Conclusions: This genomics tool offers the opportunity for large-scale biological approaches combining both expression and genomic-level sequence analysis using as a template an entire vertebrate genome. © 2004 Balciunas et al; licensee BioMed Central Ltd.
    Citation to related work
    Springer Science and Business Media LLC
    Has part
    BMC Genomics
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    ae974a485f413a2113503eed53cd6c53
    http://dx.doi.org/10.34944/dspace/5638
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