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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MicroRNA26 attenuates vascular smooth muscle maturation via endothelial BMP signalling.
TL;DR: The data suggest that miR26a modulates BMP signalling in endothelial cells and indirectly promotes a differentiated smooth muscle phenotype, highlighting how crosstalk from BMP-responsive endothelium to smooth muscle is important for smooth muscle differentiation.
Journal ArticleDOI
A mechanism for red coloration in vertebrates
Matthew B. Toomey,Cristiana I. Marques,Pedro Araújo,Delai Huang,Siqiong Zhong,Yu Liu,Gretchen D Schreiner,Connie A. Myers,Paulo Pereira,Sandra Afonso,Pedro Andrade,Małgorzata Anna Gazda,Ricardo Lopes,Ivan Viegas,Rebecca E. Koch,Maureen E. Haynes,Dustin John Smith,Yohey Ogawa,D. A. Murphy,Rachel E. Kopec,David M. Parichy,Miguel Carneiro,Joseph C. Corbo +22 more
TL;DR: In this article , it was shown that two enzymes, cytochrome P450 2J19 (CYP2J19) and 3-hydroxybutyrate dehydrogenase 1-like (BDH1L), are sufficient to catalyze this conversion.
Posted ContentDOI
Dysregulated heparan sulfate proteoglycan metabolism promotes Ewing sarcoma tumor growth
Elena A. Vasileva,Mikako Warren,Mikako Warren,Timothy J. Triche,Timothy J. Triche,James F. Amatruda,James F. Amatruda +6 more
TL;DR: In this article, a new genetic model of Ewing sarcoma was developed based on Cre-inducible expression of human EWSR1-FLI1 in wild type zebrafish, which causes rapid onset of SRBCTs at high penetrance.
Posted ContentDOI
Cellular and molecular mechanisms of photoreceptor tuning for prey capture in larval zebrafish
TL;DR: It is shown that UV-cones in the larval zebrafish area temporalis are specifically tuned for UV-bright prey capture in their upper frontal visual field, which uses the signal from a single cone at a time.
Journal ArticleDOI
Chimeric efferocytic receptors improve apoptotic cell clearance and alleviate inflammation
Sho Morioka,Daiki Kajioka,Yusuke Yamaoka,Rochelle M Ellison,Turan Tufan,Inge L. Werkman,Shinji Tanaka,Brady J. Barron,Satoshi Ito,Sarah Kucenas,Mark D. Okusa,Kodi S. Ravichandran +11 more
TL;DR: In this article , the authors fused a specific signaling domain within the cytoplasmic adapter protein ELMO1 to the extracellular phosphatidylserine recognition domains of the efferocytic receptors BAI1 or TIM4, generating BELMO and TELMO, respectively.
References
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Journal ArticleDOI
Stages of embryonic development of the zebrafish.
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
Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.
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.