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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Overexpression of Akt1 enhances adipogenesis and leads to lipoma formation in zebrafish.
Che-Yu Chu,Chi-Fang Chen,R. Samuel Rajendran,Chia-Ning Shen,Te-Hao Chen,Chueh-Chuan Yen,Chueh-Chuan Yen,Chih-Kuang Chuang,Dar-Shong Lin,Chung-Der Hsiao +9 more
TL;DR: The findings of this study provide direct evidence that Akt1 signaling plays an important role in balancing normal levels of fat tissue in vivo and could be a new powerful model to screen novel drugs for the treatment of human obesity.
Journal ArticleDOI
2C-Cas9: a versatile tool for clonal analysis of gene function
Vincenzo Di Donato,Flavia De Santis,Thomas O. Auer,Noé Testa,Héctor Sánchez-Iranzo,Nadia Mercader,Jean-Paul Concordet,Filippo Del Bene +7 more
TL;DR: It is shown that tissue-specific gene disruption can be achieved by driving Cas9 expression with the Gal4/UAS system, and this technique has the potential to be applied to diverse model organisms, enabling tissue- specific loss-of-function and phenotypic characterization of live and fixed tissues.
Journal ArticleDOI
IGFBP1 increases β-cell regeneration by promoting α- to β-cell transdifferentiation.
Jing Lu,Ka-Cheuk Liu,Nadja Schulz,Christos Karampelias,Jérémie Charbord,Agneta Hilding,Agneta Hilding,Linn Rautio,Philippe Bertolino,Claes-Göran Östenson,Claes-Göran Östenson,Kerstin Brismar,Kerstin Brismar,Olov Andersson +13 more
TL;DR: In this article, the authors performed whole-genome expression profiling of zebrafish islets and identified 11 secreted proteins that are upregulated during β-cell regeneration at supraphysiological levels.
Journal ArticleDOI
New transgenic reporters identify somatosensory neuron subtypes in larval zebrafish.
Ana Marie S. Palanca,Sung-Ling Lee,Laura Yee,Carlee Joe-Wong,Le A. Trinh,Elizabeth Hiroyasu,Majid Husain,Scott E. Fraser,Matteo Pellegrini,Alvaro Sagasti +9 more
TL;DR: Surprisingly, although these analyses revealed diversity in peripheral axon branching and central axon projection, PKCα/p2rx3a/trpA1b‐expressing RB cells did not possess obvious characteristic morphological features, suggesting that even within this molecularly defined subtype, individual neurons may possess distinct properties.
Journal ArticleDOI
Dopamine Modulates the Activity of Sensory Hair Cells
Cecilia P. Toro,Josef G. Trapani,Itallia V. Pacentine,Reo Maeda,Lavinia Sheets,Weike Mo,Teresa Nicolson +6 more
TL;DR: Dopaminergic efferent fibers innervate the acousticolateralis epithelium of the zebrafish during development but do not directly form synapses with hair cells, but a member of the D1-like receptor family, D1b, tightly localizes to ribbon synapses in inner ear and lateral-line hair cells.
References
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Journal ArticleDOI
GAL4-VP16 is an unusually potent transcriptional activator
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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.