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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ptf1a+ , ela3l- cells are developmentally maintained progenitors for exocrine regeneration following extreme loss of acinar cells in zebrafish larvae.
TL;DR: New transgenic models for exocrine pancreas regeneration in zebrafish reveal a novel potential exocrine progenitor as the source for a two-phase regeneration process after complete acinar cell loss and show a conserved requirement for Wnt signaling in exocrine tissue expansion.
Posted ContentDOI
An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish
Dianne Lumaquin,Eleanor Johns,Joshua M. Weiss,Emily Montal,Olayinka Ooladipupo,Abderhman Abuhashem,Richard M. White +6 more
TL;DR: A lipid droplets transgenic reporter is created by fusing tdTOMATO to Perilipin-2 (PLIN2), a lipid droplet structural protein, and a novel regulators of lipolysis are tested, revealing an unexpected role for nitric oxide in modulating adipocyte lipid Droplets.
Journal ArticleDOI
Cdc6 cooperates with c-Myc to promote genome instability and epithelial to mesenchymal transition (EMT) in zebrafish
Ching-Hung Chen,Dar-Shong Lin,Chieh-Wen Cheng,Chun-Ju Lin,Yu-Kang Lo,Chueh-Chuan Yen,Alan Yueh-Luen Lee,Chung-Der Hsiao +7 more
TL;DR: Co-overexpression of cdc6 and c-myc in transgenic zebrafish skin triggered tumor-like transformation, apoptosis attenuation, genomic instability, and EMT, hallmarks of malignant tumorigenesis, provide the future utility of this model for easy and non-invasive detection and for identification of new anti-cancer drug.
Journal ArticleDOI
Rac2 Regulates the Migration of T Lymphoid Progenitors to the Thymus during Zebrafish Embryogenesis.
TL;DR: Mechanistically, it is demonstrated that Rac2 regulates homing of T lymphoid progenitor through Pak1-mediated AKT pathway, which is an important function of Rac2 in directing T lymphoids progenitors migration to the thymus during zebrafish embryogenesis.
Journal ArticleDOI
Correction: Vegfc acts through ERK to induce sprouting and differentiation of trunk lymphatic progenitors.
Masahiro Shin,Ira Male,Timothy J. Beane,Jacques A. Villefranc,Fatma O. Kok,Lihua Julie Zhu,Nathan D. Lawson +6 more
TL;DR: There was an error published in Development 143 where an activated myc-tagged form of MEK was generated from zebrafish map2k2b by providing an incorrect serine residue number for the map2K2b mutation.
References
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Journal ArticleDOI
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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.