Nov 24, 2015 3dimensional spatial registration of gal4 lines. Expression levels of the reporter require that the enhancer trap insert and come under control of an endogenous enhancer element. Software for zebrafish behavioral analysis transgenic lines gal4 enhancer trap database. We also compared the tissue expression pattern of our collection with that from a previously described gal4 enhancer trap collection np of the drosophila genome resource center dgrc. Oct 01, 2016 overall, 98% stanex lines partially or fully reproduced tissuespecific expression detected previously by rnaseq.
In this collection, tol2mediated transgenesis was used to generate gal4 enhancer trap lines. Primers dashed arrows designed against either the 5. The gal4 lines can be converted into other reporters through the use of rmce, providing a versatile toolkit. In the enhancer trap vectors, the reporter gene is fused to a minimal promoter, which is unable to drive the reporter gene expression alone but can be activated by neighbouring cis. Genomic dna from a gal4 enhancer trap strain containing integrated pgawb pelement engineered to contain gal4 is digested with a frequent cutter, hpaii, and the resulting fragments are circularized by intramolecular ligation. The original database has been taken offline by imp. A 3d searchable database of transgenic zebrafish gal4 and. An enhancer trap vector bearing a modified gal4 vp16 gene was inserted randomly into the arabidopsis genome by agrobacterium mediated transformation. Cell specific activation of the gal4 vp16 gene by cellular enhancer results in activation of a linked gfp gene, allowing simple characterisation of. A 3d searchable database of transgenic zebrafish gal4 and cre lines for functional neuroanatomy studies. In this study, we present the molecular characterization and initial tissue expression analysis of nearly 100 novel stanex lexa.
Using transposon mutagenesis in drosophila, we constructed a library of 7404 protein trap and enhancer trap lines, the carnegie collection, to facilitate gene expression mapping at singlecell resolution. Feb 10, 2020 engert lab neural enhancer trap line resource. Gal4 enhancer trap lines are useful markers to tag specifc cell types and. An interscholastic network to generate lexa enhancer trap.
Jul 12, 2017 genetic access to small, reproducible sets of neurons is key to an understanding of the functional wiring of the brain. Maximum projections through 80 gal4 enhancer trap lines that were imaged, registered and averaged for this study. The database includes 80 gal4 enhancer trap lines and. Cell specific activation of the gal4vp16 gene by cellular enhancer results in activation of a linked gfp gene. Many of the imaged enhancer trap lines show intrinsic brainspecific expression. In this laboratory drosophila melanogaster is used as the model organism and a set of techniques known as enhancer trap analysis is employed to begin the experiments. It has also been adapted to study receptor chemicalbinding functions in vitro in cell culture. At 4 dpf, larvae were exposed to 10 mm metronidazole for 24 h, given 24 h to recover, and then tested for escape behavior at 6 dpf. To construct a high resolution database of enhancer trap lines, we first developed a pipeline for generating average brain representations for transgenic lines figure figure1a. Frontiers a 3d searchable database of transgenic zebrafish. Drosophila is one of the favored models systems for molecular and genetic studies for a number of reasons. Jul 25, 2005 a screen for lateral root development related enhancer trap lines. Automated highthroughput mapping of promoterenhancer. The ztrap database contains images of gfp and gal4ff expression patterns, and genomic dna sequences surrounding the integration sites of the gene trap and enhancer trap constructs.
Searches can be made by looking to insertions according to map position, expression pattern, affected gene or strain number. Here we report the generation of a new gal4 and credriver resource for. Basically, in the gal4 gfp enhancer trap system, a given tdna is introduced randomly into a host genome. Jun 12, 2008 getdb gal4 enhancer trap insertion database version 3.
A 3d searchable database of transgenic zebrafish gal4. The database also included a tool which allows users to jump onto a chromosomal armor or bands as well as inspect the. Gene trap constructs contain a splice acceptor and the gfp or gal4ff a modified version of the yeast gal4 transcription activator gene, and enhancer trap. Gal4 enhancer trap and gene trap in silkworm, bombyx mori. Blast analysis of the recovered flanking dna sequence using online databases. This neural enhancer trap line resource is a collection of gal4 expressing enhancer trap lines which have been registered to the zbrain for expression comparison and annotation. Brainbaseweb and braingazer are the two available user interfaces to brainbase data bases providing customized tools to access, visualize, explore and mine the data. A drosophila lexa enhancertrap resource for developmental. Genetic access to small, reproducible sets of neurons is key to an understanding of the functional wiring of the brain. Additional gal4 transgenic lines are described in an online database. Each panel is the average of at least three brains. Expression profiling of receptor genes reveals neurochemical segmentation of the brain. Enhancer trap lines with strong expression in the nervous system are included in our brain browser, an atlas of zebrafish transgenic lines.
Gal4gfp enhancer trap lines for genetic manipulation of lateral. Tissuespecific or ubiquitous enhancers drive expression of gal4 that results in transcriptional activation from the gal4 binding sites uas to drive expression of a gene of interest, and is widely used for expression of fluorescently tagged proteins and for gene knockdown by rnai. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with coexpression of cre and gal4. Pdf zebrafish enhancer trap transgenic line database zetrap 2.
Aug 11, 2009 subsequently, a tol2 transposon mediated selfreporting gal4 enhancer trap screen was performed to establish a collection of cell typespecific activator lines with characterized transactivation kinetics that will provide a useful resource for targeted transgene activation in zebrafish. The p element can carry a reporter gene and an eye color marker as well as various bacterial elements necessary for cloning. The database includes 80 gal4 enhancer trap lines and 9 cre. A collection of enhancer trap insertional mutants for. Genetic targeting and anatomical registration of neuronal. An example gal4 uas system, with gal4 lines and uas reporter lines. The integration sites are mapped onto the ensembl zebrafish genome by inhouse blat analysis and can be viewed on the ztrap and ensembl genome browsers. Database searches were performed using the blast program. Optimized gal4 genetics for permanent gene expression mapping. Methods for automated fluorescence imaging allow highthroughput examination of reporter expression patterns in zebrafish embryos. Zebrafish enhancer trap transgenic line database zetrap 2. Metazoan physiology depends on intricate patterns of gene expression that remain poorly known.
Mar 19, 2017 enhancer trap system is a valuable tool for identifying regulatory elements. The enhancer trap construct carries the egfp gene controlled by a partial epithelial promoter from the keratin8 gene. A gal4driver line resource for drosophila neurobiology. The gal4 uas system is a biochemical method used to study gene expression and function in organisms such as the fruit fly. The integration sites are mapped onto the ensembl zebrafish genome by inhouse blat analysis and can be viewed on the z trap and ensembl genome browsers. For example, ok107gal4 traps an enhancer of the eyeless gene and is. Show full abstract transgenic lines have been collected and compiled in the form of an online databasethe zebrafish enhancer trap lines database zetrap. Gal4 gfp enhancer trap lines for analysis of stomatal guard.
Enhancer trap an enhancer trap element is a mobile dna element that is usually a p element. The first is necessary for random insertion in the genome, the latter is necessary for identification of the spatial regulation by the enhancer. The ability to reuse the enhancer fragments greatly enhances the value of the image database we report here. Gal4gfp enhancer trap lines for genetic manipulation of. This was not unexpected as many of the lines in the haseloff gal4 gfp enhancer trap database were initially selected for in a root gfp screen laplaze et al. Mar 15, 20 gal4gfp enhancer trap system its a system for gal4 targeted gene expression. Neurochemical organization of the drosophila brain visualized. A collection of 401 gal4 gfp enhancer trap et lines generated by root transformation of wildtype arabidopsis plants ecotype c24 j haseloff and s hodge, unpublished results was screened for gfp expression in lateral root founder cells or during lateral root development.
Genomic resources for zebrafish general information zfin. They are applied to mapping promoter enhancer interactions in. To aid in navigating the brain and interpreting enhancer trap expression patterns, we also imaged. Introduction to enhancer trap analysis hofstra people. Show full abstract transgenic lines have been collected and compiled in the form of an online database the zebrafish enhancer trap lines database zetrap. A 3d searchable database of transgenic zebrafish gal4 and cre. Getdb gal4 enhancer trap insertion database version 3. Molecular identification of a root apical cellspecific and.
Pdf a 3d searchable database of transgenic zebrafish gal4. Nov 24, 2015 a 3d searchable database of transgenic zebrafish gal4 and cre lines for functional neuroanatomy studies. Another zebrafish enhancer trap database is maintained by harold burgess lab. The database includes 80 gal4 enhancer trap lines and 9 cre enhancer trap lines. This database contains information on reporter expressions and positions of transposon vectors mutators in transposon insertion lines enhancer trap lines and gene trap lines, and is integrated into silkworm genome database, kaikobase shimomura et al. Using confocal microscopy, we have imaged the cellularlevel expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 gal4 enhancer trap lines, 9 cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene. Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. An enhancer trap vector bearing a modified gal4vp16 gene was inserted randomly into the arabidopsis genome by agrobacterium mediated transformation. Prepontine nongiant neurons drive flexible escape behavior. Gathers insertion site, expression pattern and mutant phenotype from gal4 enhancer trap lines. This software also enables the intersectional expression of transgenes to be. To aid in navigating the brain and interpreting enhancer trap expression patterns, we also imaged 20 transgenic lines with genetically defined expression patterns. Average brain representations of gal4 enhancer trap lines.
The framework has been originally developed in context of vt and enhancer trap gal4 lines of adult drosophila generated by barry dickson and his group over. Prior studies have used collections of gal4expressing lines based on enhancer traps. Modified transposon dna and transposase rna were delivered into zebrafish embryos by microinjection to produce heritable insertions in the zebrafish genome. Drosophila gal4 enhancer trap characterization in an. It was developed by hitoshi kakidani and mark ptashne, and.
Getdb is an integrated database compiling insertion site, expression pattern and mutant phenotype of a large collection 6966 of gal4 enhancer. The enhancer trap construct contains a transposable element and a reporter gene. The brain browser software provides a simple way to identify enhancer trap gal4 lines that label a specific 3dimensional region of the brain. However, the number of welldefined lexa enhancer trap insertions remains limited. A drosophila lexa enhancertrap resource for developmental biology and. To assist with this, the current data set includes 20 transgenic lines that were generated with defined promoters or bac transgenesis that reproduce known neuroanatomical expression patterns of specific genes. Brain is a software framework designed to host, manage, visualize and. Video articles in jove about gal4 enhancer trap include induction and diagnosis of tumors in drosophila imaginal disc epithelia, targeting olfactory bulb neurons using combined in vivo electroporation and gal4 based enhancer trap zebrafish lines, investigating tissue and organspecific phytochrome responses using facsassisted celltype specific expression profiling in arabidopsis. Full resolution stacks can be downloaded from the website and lines can be ordered from the engert lab. An enhancer trap vector bearing a modified gal4vp16 gene was inserted randomly into the arabidopsis genome by agro bacterium mediated transformation.
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