Journal ArticleDOI
The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs.
Kristen M. Kwan,Esther Fujimoto,Clemens Grabher,Benjamin D. Mangum,Melissa Hardy,Douglas Simon Campbell,John M. Parant,H. Joseph Yost,John P. Kanki,Chi Bin Chien +9 more
TLDR
The Tol2kit greatly facilitates zebrafish transgenesis, simplifies the sharing of clones, and enables large‐scale projects testing the functions of libraries of regulatory or coding sequences.Abstract:
Transgenesis is an important tool for assessing gene function. In zebrafish, transgenesis has suffered from three problems: the labor of building complex expression constructs using conventional subcloning; low transgenesis efficiency, leading to mosaicism in transient transgenics and infrequent germline incorporation; and difficulty in identifying germline integrations unless using a fluorescent marker transgene. The Tol2kit system uses site-specific recombination-based cloning (multisite Gateway technology) to allow quick, modular assembly of [promoter]-[coding sequence]-[3' tag] constructs in a Tol2 transposon backbone. It includes a destination vector with a cmlc2:EGFP (enhanced green fluorescent protein) transgenesis marker and a variety of widely useful entry clones, including hsp70 and beta-actin promoters; cytoplasmic, nuclear, and membrane-localized fluorescent proteins; and internal ribosome entry sequence-driven EGFP cassettes for bicistronic expression. The Tol2kit greatly facilitates zebrafish transgenesis, simplifies the sharing of clones, and enables large-scale projects testing the functions of libraries of regulatory or coding sequences.read more
Citations
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Journal ArticleDOI
Fxr signaling and microbial metabolism of bile salts in the zebrafish intestine
Jia Wen,Gilberto Padilla Mercado,Alyssa Volland,Heidi L. Doden,Colin R. Lickwar,Taylor Crooks,Genta Kakiyama,Cecelia Kelly,Jordan L. Cocchiaro,Jason M. Ridlon,John F. Rawls +10 more
TL;DR: In this paper, the authors provided a cellular atlas of the zebrafish intestinal epithelium and uncovered roles for Fxr in transcriptional and differentiation programs in ileal and other cell types.
Journal ArticleDOI
Embryonic exposure to excess thyroid hormone causes thyrotrope cell death
TL;DR: Using the zebrafish embryo as a model for thyroid axis development, the authors characterized the ontogeny of negative feedback regulation of thyrotrope function and examined the effect of excess TH on thyrope development.
Journal ArticleDOI
Notch-Mediated Determination of Hair-Bundle Polarity in Mechanosensory Hair Cells of the Zebrafish Lateral Line
TL;DR: Through a combination of theoretical and experimental approaches, it is shown that Notch1a-mediated lateral inhibition produces a bistable switch that reliably gives rise to cell pairs of opposite polarity.
Book ChapterDOI
H2O2: A Chemoattractant?
Balázs Enyedi,Philipp Niethammer +1 more
TL;DR: This work presents a detailed protocol of the zebrafish tail fin wounding assay and explains how to use it for analyzing leukocyte chemotaxis in vivo, and outlines a method for H2O2 measurement in live zebra fish larvae using the genetically encoded sensor HyPer on a wide-field and a spinning disk confocal microscope.
Journal ArticleDOI
Hypothalamic Projections to the Optic Tectum in Larval Zebrafish.
Lucy A. Heap,Gilles Vanwalleghem,Andrew W. Thompson,Itia A. Favre-Bulle,Halina Rubinsztein-Dunlop,Ethan K. Scott +5 more
TL;DR: Tectal processing in larval zebrafish is modulated by hypothalamic inhibitory inputs to the deep tectal neuropil, suggesting that this role extends beyond the visual system, and beyond the processing of exclusively retinal inputs.
References
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Journal ArticleDOI
DNA Cloning Using In Vitro Site-Specific Recombination
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