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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A Novel Model for Development, Organization, and Function of Gonadotropes in Fish Pituitary
TL;DR: Two transgenic zebrafish lines with labeled gonadotropes provide a powerful tool for investigating the development, anatomy, and function of the reproductive axis in lower vertebrates.
Journal ArticleDOI
Aciculin interacts with filamin C and Xin and is essential for myofibril assembly, remodeling and maintenance.
Sibylle Molt,John B. Bührdel,Sergiy Yakovlev,Peter Schein,Zacharias Orfanos,Gregor Kirfel,Lilli Winter,Gerhard Wiche,Peter F.M. van der Ven,Wolfgang Rottbauer,Steffen Just,Alexey M. Belkin,Dieter O. Fürst +12 more
TL;DR: The identified the dystrophin-binding protein aciculin (also known as phosphoglucomutase-like protein 5, PGM5) as a new interaction partner of FLNc and Xin and points to a thus far unappreciated, but essential, function of acicul in myofibril formation, maintenance and remodeling.
Journal ArticleDOI
Etsrp/Etv2 Is Directly Regulated by Foxc1a/b in the Zebrafish Angioblast
Matthew B. Veldman,Shuo Lin +1 more
TL;DR: Mesodermal foxc1a/b is identified as a direct upstream regulator of etsrp in angioblasts, establishing a new molecular link in the process of mesoderm specification into angioblast.
Journal ArticleDOI
Dynamic analysis of BMP-responsive smad activity in live zebrafish embryos.
TL;DR: This transgenic Smad1/5/8 reporter line will be useful for determining ligand and receptor requirements for specific domains of BMP activity, as well as for genetic and pharmacological screens aimed at identifying enhancers or suppressors of canonical BMP signaling.
Journal ArticleDOI
Liver-Specific Expressions of HBx and src in the p53 Mutant Trigger Hepatocarcinogenesis in Zebrafish
Jeng-Wei Lu,Wan-Yu Yang,Su-Mei Tsai,Yueh-Min Lin,Pen-Heng Chang,Jim-Ray Chen,Jim-Ray Chen,Horng-Dar Wang,Jen-Leih Wu,Shiow-Lian Catherine Jin,Chiou-Hwa Yuh,Chiou-Hwa Yuh,Chiou-Hwa Yuh +12 more
TL;DR: It is shown that the synergistic action of HBx and p53 mutation triggers progressive hepatocellular carcinoma (HCC) formation via src activation in zebrafish, which mimics human HCC formation and provides potential in vivo platforms for drug screening for therapies for human liver cancer.
References
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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.