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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PRL3-DDX21 Transcriptional Control of Endolysosomal Genes Restricts Melanocyte Stem Cell Differentiation.
Jeanette A. Johansson,Jeanette A. Johansson,Kerrie L. Marie,Kerrie L. Marie,Kerrie L. Marie,Yuting Lu,Yuting Lu,Alessandro Brombin,Alessandro Brombin,Cristina Santoriello,Zhiqiang Zeng,Zhiqiang Zeng,Judith Zich,Judith Zich,Philippe Gautier,Alex von Kriegsheim,Hannah Brunsdon,Hannah Brunsdon,Ann P. Wheeler,Marcel Dreger,Marcel Dreger,Douglas R. Houston,Christopher M. Dooley,Christopher M. Dooley,Andrew H. Sims,Elisabeth M. Busch-Nentwich,Elisabeth M. Busch-Nentwich,Leonard I. Zon,Robert S. Illingworth,E. Elizabeth Patton,E. Elizabeth Patton +30 more
TL;DR: It is shown that PRL3 binds to the RNA helicase DDX21, thereby restricting productive transcription by RNAPII at master transcription factor (MITF)-regulated endolysosomal vesicle genes in MSC regeneration.
Journal ArticleDOI
Transgenic retinoic acid sensor lines in zebrafish indicate regions of available embryonic retinoic acid.
Amrita Mandal,Amrita Mandal,Ariel Rydeen,Ariel Rydeen,Jane Anderson,Mollie R.J. Sorrell,Tomas Zygmunt,Jesús Torres-Vázquez,Joshua S. Waxman +8 more
TL;DR: Novel transgenic RA sensors in which a RA receptor (RAR) ligand‐binding domain (RLBD) is fused to the Gal4 DNA‐ binding domain (GDBD) or a VP16‐G DBD (VPBD) construct are generated, demonstrating there may be almost ubiquitous availability of RA within the early embryo.
Journal ArticleDOI
Pax6 organizes the anterior eye segment by guiding two distinct neural crest waves
Masanari Takamiya,Johannes Stegmaier,Andrei Yu Kobitski,Benjamin Schott,Benjamin D. Weger,Dimitra Margariti,Angel R. Cereceda Delgado,Victor Gourain,Tim Scherr,Lixin Yang,Sebastian Sorge,Jens C. Otte,Volker Hartmann,Jos van Wezel,Rainer Stotzka,Thomas Reinhard,Günther Schlunck,Thomas Dickmeis,Sepand Rastegar,Ralf Mikut,Gerd Ulrich Nienhaus,Uwe Strähle +21 more
TL;DR: This work traces the origin of the ocular NC cells to two distinct NC populations that differ in the maintenance of sox10 expression, Wnt signalling, origin, route, mode and destination of migration and provides evidence that the lens whose formation is pax6a/b-dependent and lens-derived TGFβ signals contribute to the building of the anterior segment.
Journal ArticleDOI
Unique and non-redundant function of csf1r paralogues in regulation and evolution of post-embryonic development of the zebrafish.
Joana Caetano-Lopes,Joana Caetano-Lopes,Katrin Henke,Katrin Henke,Katia Urso,Katia Urso,Jeffrey Duryea,Julia F. Charles,Julia F. Charles,Matthew L. Warman,Matthew L. Warman,Matthew P. Harris,Matthew P. Harris +12 more
TL;DR: The sub-functionalization of csf1r paralogous genes in teleost fish coincides with the evolution of a novel function in regulating pigmentation, which is not observed in mammals or birds.
Journal ArticleDOI
Nifurpirinol: A more potent and reliable substrate compared to metronidazole for nitroreductase‐mediated cell ablations
David Bergemann,Laura Massoz,Jordane Bourdouxhe,Claudio Andrès Carril Pardo,Marianne Voz,Bernard Peers,Isabelle Manfroid +6 more
TL;DR: It is shown that nifurpirinol induces robust and reliable ablations at concentrations 2,000 fold lower than metronidazole and three times below its own toxic concentration, and suggested that its use could be extended to many other cell types, especially if difficult to ablate, or when combined pharmacological treatments are desired.
References
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Journal ArticleDOI
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TL;DR: A series of stages for development of the embryo of the zebrafish, Danio (Brachydanio) rerio is described, providing for flexibility and continued evolution of the staging series as the authors learn more about development in this species.
Journal ArticleDOI
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Journal ArticleDOI
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Sean Munro,Hugh R.B. Pelham +1 more
TL;DR: A cDNA clone is characterized that encodes a protein related to the 70 kd heat shock protein, but is expressed in normal rat liver, and it is identical with two previously described proteins: GRP78, whose synthesis is induced by glucose starvation, and BiP, which is found bound to immunoglobulin heavy chains in pre-B 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
DNA Cloning Using In Vitro Site-Specific Recombination
TL;DR: A method called recombinational cloning is described that uses in vitro site-specific recombination to accomplish the directional cloning of PCR products and the subsequent automatic subcloning of the DNA segment into new vector backbones at high efficiency.