A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations
TLDR
It is shown that TrpR/tUAS transgenes are not silenced in subsequent generations of zebrafish, which is a major improvement over some of the existing systems, such as Gal4/gUAS and the Q-system.Abstract:
The ability to visualize and manipulate cell fate and gene expression in specific cell populations has made gene expression systems valuable tools in developmental biology studies. Here, we describe a new system that uses the E. coli tryptophan repressor and its upstream activation sequence (TrpR/tUAS) to drive gene expression in stable zebrafish transgenic lines and in mammalian cells. We show that TrpR/tUAS transgenes are not silenced in subsequent generations of zebrafish, which is a major improvement over some of the existing systems, such as Gal4/gUAS and the Q-system. TrpR transcriptional activity can be tuned by mutations in its DNA-binding domain, or silenced by Gal80 when fused to the Gal4 activation domain. In cases in which more than one cell population needs to be manipulated, TrpR/tUAS can be used in combination with other, existing systems.read more
Citations
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Binary systemforregulating transgene expression inmice: Targeting int-2 geneexpression withyeast GAL4/UAS control elements
TL;DR: In this paper, a binary transgenetic system consisting of two types of transgenic mice, targets and transactivators, was proposed to deal with the problem of genetic cross-crossing.
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Fovea-like Photoreceptor Specializations Underlie Single UV Cone Driven Prey-Capture Behavior in Zebrafish.
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CRISPR/Cas9-mediated conversion of eGFP - into Gal4 -transgenic lines in zebrafish
TL;DR: This conversion allows the in-depth analysis of the former eGFP-expressing cell population; with the Gal4-upstream activating sequence (UAS) system, diverse UAS transgenes can be transactivated.
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Remote z-scanning with a macroscopic voice coil motor for fast 3D multiphoton laser scanning microscopy
TL;DR: A focusing technique based on a remote movable mirror that is conjugate to the specimen plane and translated by a voice coil motor is developed and used for 3D multiphoton calcium imaging in the adult zebrafish brain.
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Transgenic fish systems and their application in ecotoxicology
TL;DR: Progress in the development and application of transgenic fish (principally zebrafish) for studies in environmental toxicology is assessed and a viewpoint on future development opportunities is provided.
References
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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.
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TL;DR: 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.
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EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion
Jörg Wiedenmann,Sergey Ivanchenko,Franz Oswald,Florian Schmitt,Carlheinz Röcker,Anya Salih,Klaus-Dieter Spindler,G. Ulrich Nienhaus +7 more
TL;DR: A gene encoding a fluorescent protein from the stony coral Lobophyllia hemprichii has been cloned in Escherichia coli and characterized by biochemical and biophysical methods and is found to be tetrameric, with strong Forster resonance coupling among the individual fluorophores.
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Negative effect of the transcriptional activator GAL4
Grace Gill,Mark Ptashne +1 more
TL;DR: It is suggested that this inhibition, which the authors call squelching, reflects titration of a transcription factor by the activating region of GAL4, and that more efficient activators inhibit more strongly and that inhibition does not depend on the DNA-binding domain.
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Bright monomeric red fluorescent protein with an extended fluorescence lifetime
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TL;DR: TagRFP is reported, a monomeric red fluorescent protein, which is characterized by high brightness, complete chromophore maturation, prolonged fluorescence lifetime and high pH-stability, which makes it an excellent tag for protein localization studies and fluorescence resonance energy transfer (FRET) applications.