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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The Warburg effect is necessary to promote glycosylation in the blastema during zebrafish tail regeneration.
Jason Sinclair,David R. Hoying,Erica Bresciani,Damian Dalle Nogare,Carli D. Needle,Alexandra Berger,Weiwei Wu,Kevin Bishop,Abdel G. Elkahloun,Ajay B. Chitnis,Paul P. Liu,Shawn M. Burgess +11 more
TL;DR: In this article, the authors used a larval tail regeneration assay in the zebrafish Danio rerio, which serves as an ideal model of appendage regeneration due to its easy manipulation, relatively simple mixture of cell types, and superior imaging properties.
Journal ArticleDOI
Characterization of Ribeye Subunits in Zebrafish Hair Cells Reveals That Exogenous Ribeye B-Domain and CtBP1 Localize to the Basal Ends of Synaptic Ribbons
TL;DR: How exogenously expressed Ribeye domains interact with synaptic ribbons in vivo is revealed, and a potential organization of elements within the ribbon body is suggested.
Journal ArticleDOI
A transgenic zebrafish model expressing KIT‐D816V recapitulates features of aggressive systemic mastocytosis
Tugce B. Balci,Sergey V. Prykhozhij,Evelyn M. Teh,Sahar I. Da'as,Eileen R. McBride,Eileen R. McBride,Robert S. Liwski,Ian C. Chute,Daniel Leger,Stephen M. Lewis,Jason N. Berman +10 more
TL;DR: This study represents the first zebrafish model of a mast cell disease with an aggressive adult phenotype and embryonic markers that could be exploited to screen for novel agents in SM.
Hand2 elevates cardiomyocyte production during zebrafish heart development and regeneration
Yocheved L. Schindler,Kristina M. Garske,Jinhu Wang,Beth A. Firulli,Anthony B. Firulli,Kenneth D. Poss,Deborah Yelon +6 more
TL;DR: It is found that overexpression of hand2 in the early zebrafish embryo is able to enhance cardiomyocyte production, resulting in an enlarged heart with a striking increase in the size of the outflow tract and this results contribute to the understanding of the potential origins of congenital heart disease.
Journal ArticleDOI
Enhancing regeneration after acute kidney injury by promoting cellular dedifferentiation in zebrafish
Lauren Brilli Skvarca,Hwa In Han,Eugenel B. Espiritu,Maria A. Missinato,Maria A. Missinato,Elizabeth R. Rochon,Michael D. McDaniels,Abha S. Bais,Beth L. Roman,Joshua S. Waxman,Simon C. Watkins,Alan J. Davidson,Michael Tsang,Neil A. Hukriede +13 more
TL;DR: In this paper, the regenerative mechanisms of PTBA were investigated in zebrafish models of larval renal injury and adult cardiac injury, and it was shown that PTBA using an esterified prodrug (UPHD25) increases the reactivation of the renal progenitor gene Pax2a, enhances dedifferentiation of RTECs, reduces Kidney injury molecule-1 (Kim-1) expression, and lowers the number of infiltrating macrophages.
References
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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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Nathan C. Shaner,Robert E. Campbell,Robert E. Campbell,Paul Steinbach,Ben N G Giepmans,Amy E. Palmer,Roger Y. Tsien +6 more
TL;DR: The latest red version matures more completely, is more tolerant of N-terminal fusions and is over tenfold more photostable than mRFP1, and three monomers with distinguishable hues from yellow-orange to red-orange have higher quantum efficiencies.
Journal ArticleDOI
An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein.
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.