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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Journal ArticleDOI
Unlimited Genetic Switches for Cell-Type-Specific Manipulation.
TL;DR: CaSSA is a new genetic tool that conceptually works as an unlimited number of recombinases and will facilitate genetic access to cell types in diverse organisms.
Posted ContentDOI
Wnt/β-catenin regulates an ancient signaling network during zebrafish scale development
TL;DR: These analyses demonstrate the utility of zebrafish scale development as a tractable system in which to elucidate mechanisms of developmental patterning and morphogenesis, and suggest a single, ancient origin of skin appendage patterning mechanisms in vertebrates.
Journal ArticleDOI
Tbx20 drives cardiac progenitor formation and cardiomyocyte proliferation in zebrafish
TL;DR: A previously unappreciated role for Tbx20 in promoting cardiac progenitor formation in vertebrates is highlighted and a novel function for its activation domain in cardiac cell proliferation during embryogenesis is revealed.
Journal ArticleDOI
Wild-type cone photoreceptors persist despite neighboring mutant cone degeneration.
TL;DR: The zebrafish is a cone-rich animal model in which the potential for dying cones to kill neighboring healthy cones can be evaluated and it is found that the death of inoperative cones does not cause neighboring WT cone loss.
Journal ArticleDOI
Novel cardiovascular gene functions revealed via systematic phenotype prediction in zebrafish
Gabriel Musso,Gabriel Musso,Murat Tasan,Murat Tasan,Christian Mosimann,Christian Mosimann,John E. Beaver,Eva Plovie,Logan A. Carr,Logan A. Carr,Hon Nian Chua,Julie Dunham,Khalid Zuberi,Harold Rodriguez,Quaid Morris,Leonard I. Zon,Leonard I. Zon,Frederick P. Roth,Calum A. MacRae,Calum A. MacRae +19 more
TL;DR: It is shown that systematic prioritization in zebrafish can accelerate the pace of developmental gene function discovery, and identified tmem88a, a recently described attenuator of Wnt signaling, as a discrete regulator of the patterning of intercellular coupling in the zebra fish cardiac epithelium.
References
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Sean Munro,Hugh R.B. Pelham +1 more
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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.