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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FGF1 Mediates Overnutrition-Induced Compensatory β-Cell Differentiation
TL;DR: It is demonstrated that fgf1 is necessary and FGF1 expression in β-cells is sufficient for the compensatory β-cell differentiation, and that endoplasmic reticulum stress acts upstream of FGF 1 release, suggesting that the recently discovered antidiabetes function of F GF1 may act partially through increasing β- cell differentiation.
Journal ArticleDOI
Specific connectivity between photoreceptors and horizontal cells in the zebrafish retina
Lauw J. Klaassen,Wim de Graaff,Jorrit B. van Asselt,Jan Klooster,Maarten Kamermans,Maarten Kamermans +5 more
TL;DR: The functional and morphological connectivity between various horizontal cell (HC) types (H1, H2, H3, and H4) and photoreceptors was studied in zebrafish retina and the spectral sensitivities of the HC layers were determined.
Journal ArticleDOI
Linking Virus Discovery to Immune Responses Visualized during Zebrafish Infections
TL;DR: A naturally occurring picornavirus is described that elicits strong antiviral responses in zebrafish and provides new strategies for simultaneously discovering viruses and their impact on vertebrate hosts.
Journal ArticleDOI
Poised Regeneration of Zebrafish Melanocytes Involves Direct Differentiation and Concurrent Replenishment of Tissue-Resident Progenitor Cells.
TL;DR: Tissue-resident progenitor cells that mediate regeneration of zebrafish stripe melanocytes are identified and how these cells reconstitute pigmentation is defined to serve as a means to rapidly repair injured tissue while preserving the capacity to regenerate.
Journal ArticleDOI
Cooperation between melanoma cell states promotes metastasis through heterotypic cluster formation
Nathaniel R. Campbell,Anjali Rao,Miranda V. Hunter,Magdalena Katarzyna Sznurkowska,Luzia Briker,Maomao Zhang,Maayan Baron,Silja Heilmann,Maxime Deforet,Colin Kenny,Lorenza P. Ferretti,Ting-Hsiang Huang,Sarah C. Perlee,Manik Garg,Jérémie Nsengimana,Massimo Saini,Emily Montal,Mohita Tagore,Julia Newton-Bishop,Mark R. Middleton,Pippa Corrie,David J. Adams,Roy Rabbie,Roy Rabbie,Nicola Aceto,Mitchell P. Levesque,Robert A. Cornell,Itai Yanai,Joao B. Xavier,Richard M. White +29 more
TL;DR: In this article, the TFAP2 neural crest transcription factor was identified as a master regulator of cell-cell clustering and metastasis in metastatic melanoma patients, and the authors showed that the TFAPE2 neural stem transcription factor is a key player in the formation of heterotypic clusters.
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.