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dc.creatorBalciuniene, J
dc.creatorNagelberg, D
dc.creatorWalsh, KT
dc.creatorCamerota, D
dc.creatorGeorlette, D
dc.creatorBiemar, F
dc.creatorBellipanni, G
dc.creatorBalciunas, D
dc.date.accessioned2021-01-31T18:40:00Z
dc.date.available2021-01-31T18:40:00Z
dc.date.issued2013-09-14
dc.identifier.issn1471-2164
dc.identifier.issn1471-2164
dc.identifier.doihttp://dx.doi.org/10.34944/dspace/5352
dc.identifier.other24034702 (pubmed)
dc.identifier.urihttp://hdl.handle.net/20.500.12613/5370
dc.description.abstractBackground: External development and optical transparency of embryos make zebrafish exceptionally suitable for in vivo insertional mutagenesis using fluorescent proteins to visualize expression patterns of mutated genes. Recently developed Gene Breaking Transposon (GBT) vectors greatly improve the fidelity and mutagenicity of transposon-based gene trap vectors.Results: We constructed and tested a bipartite GBT vector with Gal4-VP16 as the primary gene trap reporter. Our vector also contains a UAS:eGFP cassette for direct detection of gene trap events by fluorescence. To confirm gene trap events, we generated a UAS:mRFP tester line. We screened 270 potential founders and established 41 gene trap lines. Three of our gene trap alleles display homozygous lethal phenotypes ranging from embryonic to late larval: nsf tpl6, atp1a3atpl10 and flrtpl19. Our gene trap cassette is flanked by direct loxP sites, which enabled us to successfully revert nsf tpl6, atp1a3atpl10 and flrtpl19 gene trap alleles by injection of Cre mRNA. The UAS:eGFP cassette is flanked by direct FRT sites. It can be readily removed by injection of Flp mRNA for use of our gene trap alleles with other tissue-specific GFP-marked lines. The Gal4-VP16 component of our vector provides two important advantages over other GBT vectors. The first is increased sensitivity, which enabled us to detect previously unnoticed expression of nsf in the pancreas. The second advantage is that all our gene trap lines, including integrations into non-essential genes, can be used as highly specific Gal4 drivers for expression of other transgenes under the control of Gal4 UAS.Conclusions: The Gal4-containing bipartite Gene Breaking Transposon vector presented here retains high specificity for integrations into genes, high mutagenicity and revertibility by Cre. These features, together with utility as highly specific Gal4 drivers, make gene trap mutants presented here especially useful to the research community. © 2013 Balciuniene et al.; licensee BioMed Central Ltd.
dc.format.extent619-619
dc.language.isoen
dc.relation.haspartBMC Genomics
dc.relation.isreferencedbySpringer Science and Business Media LLC
dc.rightsCC BY
dc.rights.urihttp://creativecommons.org/licenses/by/2.0
dc.subjectZebrafish
dc.subjectInsertional mutagenesis
dc.subjectGene trap
dc.subjectGal4
dc.subjectTol2
dc.subjectnsfa
dc.subjectFleer
dc.subjectatp1a3a
dc.subjectbbs7
dc.titleEfficient disruption of Zebrafish genes using a Gal4-containing gene trap
dc.typeArticle
dc.type.genreJournal Article
dc.relation.doi10.1186/1471-2164-14-619
dc.ada.noteFor Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
dc.creator.orcidBalciunas, Darius|0000-0003-1938-3243
dc.date.updated2021-01-31T18:39:56Z
refterms.dateFOA2021-01-31T18:40:00Z


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