Journal ArticleDOI
Targeted gene expression by the Gal4‐UAS system in zebrafish
Kazuhide Asakawa,Koichi Kawakami +1 more
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TLDR
The Gal4 gene trap and enhancer trap approaches together with various UAS lines should be important tools for investigating roles of genes and cells in vertebrate systems for a long time.Abstract:
Targeted gene expression by the Gal4-UAS system is a powerful methodology for analyzing function of genes and cells in vivo and has been extensively used in genetic studies in Drosophila. On the other hand, the Gal4-UAS system had not been applied effectively to vertebrate systems for a long time mainly due to the lack of an efficient transgenesis method. Recently, a highly efficient transgenesis method using the medaka fish Tol2 transposable element was developed in zebrafish. Taking advantage of the Tol2 transposon system, we and other groups developed the Gal4 gene trap and enhancer trap methods and established various transgenic fish expressing Gal4 in specific cells. By crossing such Gal4 lines with transgenic fish lines harboring various reporter genes and effector genes downstream of UAS (upstream activating sequence), specific cells can be visualized and manipulated in vivo by targeted gene expression. Thus, the Gal4 gene trap and enhancer trap approaches together with various UAS lines should be important tools for investigating roles of genes and cells in vertebrates.read more
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An optimized fluorescent probe for visualizing glutamate neurotransmission
Jonathan S. Marvin,Bart G. Borghuis,Bart G. Borghuis,Lin Tian,Lin Tian,Joseph Cichon,Mark T. Harnett,Jasper Akerboom,Andrew Gordus,Sabine L. Renninger,Tsai Wen Chen,Cornelia I. Bargmann,Michael B. Orger,Eric R. Schreiter,Jonathan B. Demb,Wen-Biao Gan,S. Andrew Hires,Loren L. Looger +17 more
TL;DR: An intensity-based glutamate-sensing fluorescent reporter with signal-to-noise ratio and kinetics appropriate for in vivo imaging is described and its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems is validated.
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MicroRNA sponges: Progress and possibilities
TL;DR: The ways sponge and related constructs can be optimized are discussed and recent applications of this method are reviewed with particular emphasis on stable expression in cancer studies and in transgenic animals.
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Optical control of zebrafish behavior with halorhodopsin
TL;DR: Together this “optogenetic toolkit” allows loss-of-function and gain- of-function analyses of neural circuitry at high spatial and temporal resolution in a behaving vertebrate.
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Transgenic microRNA inhibition with spatiotemporal specificity in intact organisms
TL;DR: The Drosophila melanogaster microRNA sponge (miR-SP) is introduced as a powerful transgenic technology to dissect the function of every microRNA with precise spatiotemporal resolution and identified an essential role of the conserved microRNA miR-8, in neuromuscular junction formation.
Journal ArticleDOI
Tip cell-specific requirement for an atypical Gpr124- and Reck-dependent Wnt/β-catenin pathway during brain angiogenesis.
Benoit Vanhollebeke,Oliver A. Stone,Naguissa Bostaille,Chris Cho,Yulian Zhou,Emilie Maquet,Anne Gauquier,Pauline Cabochette,Shigetomo Fukuhara,Naoki Mochizuki,Jeremy Nathans,Didier Yr Dy Stainier +11 more
TL;DR: It is shown that mosaic restoration of single wild-type tip cells in Wnt/β-catenin-deficient perineural vessels is sufficient to initiate the formation of CNS vessels and provides evidence for organ-specific control of vascular invasion through tight modulation of tip cell function.
References
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Journal ArticleDOI
Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.
Andrea H. Brand,Norbert Perrimon +1 more
TL;DR: The GAL4 system, a system for targeted gene expression that allows the selective activation of any cloned gene in a wide variety of tissue- and cell-specific patterns, has been designed and used to expand the domain of embryonic expression of the homeobox protein even-skipped.
Journal ArticleDOI
Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin
Giampietro Schiavo,Fabio Benfenati,Bernard Poulain,Ornella Rossetto,Patrizia Polverino de Laureto,Bibhuti R. DasGupta,Cesare Montecucco +6 more
TL;DR: The results indicate that tetanus and botulinum B neurotoxins block neurotransmitter release by cleaving synaptobrevin-2, a protein that, on the basis of the results, seems to play a key part in neurotransmitterRelease.
Journal ArticleDOI
Molecular Reconstruction of Sleeping Beauty, a Tc1-like Transposon from Fish, and Its Transposition in Human Cells
TL;DR: Sleeping Beauty is an active DNA-transposon system from vertebrates for genetic transformation and insertional mutagenesis, and it mediates precise cut-and-paste transposition in fish as well as in mouse and human cells.
Journal ArticleDOI
GAL4-VP16 is an unusually potent transcriptional activator
TL;DR: It is shown that the hybrid protein (GAL4-VP16) activates transcription unusually efficiently in mammalian cells when bound close to, or at large distances from the gene, and suggested that the activating region of VP16 may be near-maximally potent.
Journal ArticleDOI
An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein
TL;DR: A gene encoding a fluorescent protein from a stony coral, Trachyphyllia geoffroyi, which emits green, yellow, and red light, is cloned, finding that the green-red conversion is highly sensitive to irradiation with UV or violet light, which excites the protonated form of the chromophore.