ArticleBMC genomics2013
Efficient disruption of Zebrafish genes using a Gal4-containing gene trap.
Article in BMC genomics, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 40 citations in OpenAlex.
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- Conditional mutagenesis by oligonucleotide-mediated integration of loxP sites in zebrafish.PLoS genetics · 2018Article
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- Analysis of a conditional gene trap reveals that tbx5a is required for heart regeneration in zebrafish.PloS one · 2018Article
- 2C-Cas9: a versatile tool for clonal analysis of gene function.Genome research · 2016Article
- Etv2 and fli1b function together as key regulators of vasculogenesis and angiogenesis.Arteriosclerosis, thrombosis, and vascular biology · 2015Article
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Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
Abstract
backgroundExternal 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.
resultsWe 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), atp1a3a(tpl10) and flr(tpl19). Our gene trap cassette is flanked by direct loxP sites, which enabled us to successfully revert nsf( tpl6), atp1a3a(tpl10) and flr(tpl19) 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.
conclusionsThe 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.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.